PMC:7321036 / 86645-88377 JSONTXT

Annnotations TAB JSON ListView MergeView

    LitCovid-PMC-OGER-BB

    {"project":"LitCovid-PMC-OGER-BB","denotations":[{"id":"T189","span":{"begin":1475,"end":1498},"obj":"NCBITaxon:10239"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}

    LitCovid-PD-FMA-UBERON

    {"project":"LitCovid-PD-FMA-UBERON","denotations":[{"id":"T626","span":{"begin":7,"end":12},"obj":"Body_part"},{"id":"T627","span":{"begin":120,"end":125},"obj":"Body_part"},{"id":"T628","span":{"begin":194,"end":199},"obj":"Body_part"},{"id":"T629","span":{"begin":388,"end":392},"obj":"Body_part"},{"id":"T630","span":{"begin":393,"end":398},"obj":"Body_part"},{"id":"T631","span":{"begin":519,"end":526},"obj":"Body_part"},{"id":"T632","span":{"begin":589,"end":596},"obj":"Body_part"},{"id":"T633","span":{"begin":686,"end":691},"obj":"Body_part"},{"id":"T634","span":{"begin":1201,"end":1206},"obj":"Body_part"},{"id":"T635","span":{"begin":1422,"end":1429},"obj":"Body_part"},{"id":"T636","span":{"begin":1495,"end":1500},"obj":"Body_part"},{"id":"T637","span":{"begin":1534,"end":1541},"obj":"Body_part"},{"id":"T638","span":{"begin":1702,"end":1707},"obj":"Body_part"}],"attributes":[{"id":"A626","pred":"fma_id","subj":"T626","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A627","pred":"fma_id","subj":"T627","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A628","pred":"fma_id","subj":"T628","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A629","pred":"fma_id","subj":"T629","obj":"http://purl.org/sig/ont/fma/fma24984"},{"id":"A630","pred":"fma_id","subj":"T630","obj":"http://purl.org/sig/ont/fma/fma63083"},{"id":"A631","pred":"fma_id","subj":"T631","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A632","pred":"fma_id","subj":"T632","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A633","pred":"fma_id","subj":"T633","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A634","pred":"fma_id","subj":"T634","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A635","pred":"fma_id","subj":"T635","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A636","pred":"fma_id","subj":"T636","obj":"http://purl.org/sig/ont/fma/fma68646"},{"id":"A637","pred":"fma_id","subj":"T637","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A638","pred":"fma_id","subj":"T638","obj":"http://purl.org/sig/ont/fma/fma67843"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}

    LitCovid-PD-UBERON

    {"project":"LitCovid-PD-UBERON","denotations":[{"id":"T27","span":{"begin":393,"end":398},"obj":"Body_part"},{"id":"T28","span":{"begin":1715,"end":1718},"obj":"Body_part"}],"attributes":[{"id":"A27","pred":"uberon_id","subj":"T27","obj":"http://purl.obolibrary.org/obo/UBERON_0001977"},{"id":"A28","pred":"uberon_id","subj":"T28","obj":"http://purl.obolibrary.org/obo/UBERON_2001840"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}

    LitCovid-PD-MONDO

    {"project":"LitCovid-PD-MONDO","denotations":[{"id":"T350","span":{"begin":59,"end":67},"obj":"Disease"},{"id":"T351","span":{"begin":184,"end":193},"obj":"Disease"},{"id":"T352","span":{"begin":497,"end":505},"obj":"Disease"},{"id":"T353","span":{"begin":563,"end":571},"obj":"Disease"},{"id":"T354","span":{"begin":1468,"end":1476},"obj":"Disease"}],"attributes":[{"id":"A350","pred":"mondo_id","subj":"T350","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A351","pred":"mondo_id","subj":"T351","obj":"http://purl.obolibrary.org/obo/MONDO_0005550"},{"id":"A352","pred":"mondo_id","subj":"T352","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A353","pred":"mondo_id","subj":"T353","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"},{"id":"A354","pred":"mondo_id","subj":"T354","obj":"http://purl.obolibrary.org/obo/MONDO_0005091"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}

    LitCovid-PD-CLO

    {"project":"LitCovid-PD-CLO","denotations":[{"id":"T1197","span":{"begin":0,"end":6},"obj":"http://purl.obolibrary.org/obo/CLO_0002172"},{"id":"T1198","span":{"begin":0,"end":6},"obj":"http://purl.obolibrary.org/obo/CLO_0051943"},{"id":"T1199","span":{"begin":0,"end":6},"obj":"http://purl.obolibrary.org/obo/CLO_0051958"},{"id":"T1200","span":{"begin":0,"end":6},"obj":"http://purl.obolibrary.org/obo/CLO_0051960"},{"id":"T1201","span":{"begin":7,"end":12},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T1202","span":{"begin":112,"end":125},"obj":"http://purl.obolibrary.org/obo/CLO_0051719"},{"id":"T1203","span":{"begin":194,"end":199},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T1204","span":{"begin":388,"end":392},"obj":"http://www.ebi.ac.uk/efo/EFO_0003051"},{"id":"T1205","span":{"begin":506,"end":514},"obj":"http://purl.obolibrary.org/obo/UBERON_0000158"},{"id":"T1206","span":{"begin":686,"end":691},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T1207","span":{"begin":718,"end":725},"obj":"http://purl.obolibrary.org/obo/CLO_0007225"},{"id":"T1208","span":{"begin":726,"end":730},"obj":"http://purl.obolibrary.org/obo/NCBITaxon_9925"},{"id":"T1209","span":{"begin":775,"end":782},"obj":"http://purl.obolibrary.org/obo/CLO_0007225"},{"id":"T1210","span":{"begin":783,"end":787},"obj":"http://purl.obolibrary.org/obo/NCBITaxon_9925"},{"id":"T1211","span":{"begin":793,"end":798},"obj":"http://purl.obolibrary.org/obo/CLO_0007836"},{"id":"T1212","span":{"begin":867,"end":874},"obj":"http://purl.obolibrary.org/obo/CLO_0007225"},{"id":"T1213","span":{"begin":974,"end":975},"obj":"http://purl.obolibrary.org/obo/CLO_0001020"},{"id":"T1214","span":{"begin":1040,"end":1041},"obj":"http://purl.obolibrary.org/obo/CLO_0001020"},{"id":"T1215","span":{"begin":1058,"end":1067},"obj":"http://purl.obolibrary.org/obo/BFO_0000030"},{"id":"T1216","span":{"begin":1201,"end":1206},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T1217","span":{"begin":1488,"end":1494},"obj":"http://purl.obolibrary.org/obo/CLO_0002172"},{"id":"T1218","span":{"begin":1488,"end":1494},"obj":"http://purl.obolibrary.org/obo/CLO_0051943"},{"id":"T1219","span":{"begin":1488,"end":1494},"obj":"http://purl.obolibrary.org/obo/CLO_0051958"},{"id":"T1220","span":{"begin":1488,"end":1494},"obj":"http://purl.obolibrary.org/obo/CLO_0051960"},{"id":"T1221","span":{"begin":1495,"end":1500},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}

    LitCovid-PD-CHEBI

    {"project":"LitCovid-PD-CHEBI","denotations":[{"id":"T470","span":{"begin":322,"end":334},"obj":"Chemical"},{"id":"T471","span":{"begin":519,"end":526},"obj":"Chemical"},{"id":"T472","span":{"begin":589,"end":596},"obj":"Chemical"},{"id":"T473","span":{"begin":875,"end":885},"obj":"Chemical"},{"id":"T474","span":{"begin":1051,"end":1053},"obj":"Chemical"},{"id":"T475","span":{"begin":1413,"end":1415},"obj":"Chemical"},{"id":"T476","span":{"begin":1422,"end":1429},"obj":"Chemical"},{"id":"T477","span":{"begin":1534,"end":1541},"obj":"Chemical"}],"attributes":[{"id":"A470","pred":"chebi_id","subj":"T470","obj":"http://purl.obolibrary.org/obo/CHEBI_9750"},{"id":"A471","pred":"chebi_id","subj":"T471","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"},{"id":"A472","pred":"chebi_id","subj":"T472","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"},{"id":"A473","pred":"chebi_id","subj":"T473","obj":"http://purl.obolibrary.org/obo/CHEBI_8040"},{"id":"A474","pred":"chebi_id","subj":"T474","obj":"http://purl.obolibrary.org/obo/CHEBI_33696"},{"id":"A475","pred":"chebi_id","subj":"T475","obj":"http://purl.obolibrary.org/obo/CHEBI_74067"},{"id":"A476","pred":"chebi_id","subj":"T476","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"},{"id":"A477","pred":"chebi_id","subj":"T477","obj":"http://purl.obolibrary.org/obo/CHEBI_36080"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}

    LitCovid-PubTator

    {"project":"LitCovid-PubTator","denotations":[{"id":"1929","span":{"begin":82,"end":87},"obj":"Gene"},{"id":"1930","span":{"begin":467,"end":471},"obj":"Gene"},{"id":"1931","span":{"begin":1420,"end":1421},"obj":"Gene"},{"id":"1932","span":{"begin":59,"end":69},"obj":"Species"},{"id":"1933","span":{"begin":497,"end":505},"obj":"Species"},{"id":"1934","span":{"begin":563,"end":571},"obj":"Species"},{"id":"1935","span":{"begin":793,"end":798},"obj":"Species"},{"id":"1936","span":{"begin":217,"end":220},"obj":"Chemical"},{"id":"1937","span":{"begin":237,"end":253},"obj":"Chemical"},{"id":"1938","span":{"begin":257,"end":260},"obj":"Chemical"},{"id":"1939","span":{"begin":322,"end":334},"obj":"Chemical"},{"id":"1940","span":{"begin":338,"end":341},"obj":"Chemical"},{"id":"1941","span":{"begin":402,"end":405},"obj":"Chemical"},{"id":"1942","span":{"begin":681,"end":684},"obj":"Chemical"},{"id":"1943","span":{"begin":712,"end":717},"obj":"Chemical"},{"id":"1944","span":{"begin":861,"end":866},"obj":"Chemical"},{"id":"1945","span":{"begin":875,"end":885},"obj":"Chemical"},{"id":"1946","span":{"begin":45,"end":53},"obj":"Disease"},{"id":"1947","span":{"begin":184,"end":193},"obj":"Disease"},{"id":"1948","span":{"begin":1468,"end":1487},"obj":"Disease"},{"id":"1949","span":{"begin":0,"end":6},"obj":"CellLine"},{"id":"1950","span":{"begin":117,"end":119},"obj":"CellLine"},{"id":"1951","span":{"begin":1488,"end":1494},"obj":"CellLine"}],"attributes":[{"id":"A1929","pred":"tao:has_database_id","subj":"1929","obj":"Gene:3837"},{"id":"A1930","pred":"tao:has_database_id","subj":"1930","obj":"Gene:1459"},{"id":"A1931","pred":"tao:has_database_id","subj":"1931","obj":"Gene:43740575"},{"id":"A1932","pred":"tao:has_database_id","subj":"1932","obj":"Tax:2697049"},{"id":"A1933","pred":"tao:has_database_id","subj":"1933","obj":"Tax:694009"},{"id":"A1934","pred":"tao:has_database_id","subj":"1934","obj":"Tax:694009"},{"id":"A1935","pred":"tao:has_database_id","subj":"1935","obj":"Tax:10090"},{"id":"A1936","pred":"tao:has_database_id","subj":"1936","obj":"MESH:D007854"},{"id":"A1937","pred":"tao:has_database_id","subj":"1937","obj":"MESH:C003043"},{"id":"A1938","pred":"tao:has_database_id","subj":"1938","obj":"MESH:D007854"},{"id":"A1939","pred":"tao:has_database_id","subj":"1939","obj":"MESH:D017830"},{"id":"A1940","pred":"tao:has_database_id","subj":"1940","obj":"MESH:D007854"},{"id":"A1941","pred":"tao:has_database_id","subj":"1941","obj":"MESH:D007854"},{"id":"A1942","pred":"tao:has_database_id","subj":"1942","obj":"MESH:D007854"},{"id":"A1945","pred":"tao:has_database_id","subj":"1945","obj":"MESH:D010590"},{"id":"A1946","pred":"tao:has_database_id","subj":"1946","obj":"MESH:D007239"},{"id":"A1947","pred":"tao:has_database_id","subj":"1947","obj":"MESH:D007239"},{"id":"A1948","pred":"tao:has_database_id","subj":"1948","obj":"MESH:C000657245"},{"id":"A1949","pred":"tao:has_database_id","subj":"1949","obj":"CVCL:0025"},{"id":"A1950","pred":"tao:has_database_id","subj":"1950","obj":"CVCL:4582"},{"id":"A1951","pred":"tao:has_database_id","subj":"1951","obj":"CVCL:0025"}],"namespaces":[{"prefix":"Tax","uri":"https://www.ncbi.nlm.nih.gov/taxonomy/"},{"prefix":"MESH","uri":"https://id.nlm.nih.gov/mesh/"},{"prefix":"Gene","uri":"https://www.ncbi.nlm.nih.gov/gene/"},{"prefix":"CVCL","uri":"https://web.expasy.org/cellosaurus/CVCL_"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}

    LitCovid-sentences

    {"project":"LitCovid-sentences","denotations":[{"id":"T776","span":{"begin":0,"end":166},"obj":"Sentence"},{"id":"T777","span":{"begin":167,"end":423},"obj":"Sentence"},{"id":"T778","span":{"begin":424,"end":661},"obj":"Sentence"},{"id":"T779","span":{"begin":662,"end":932},"obj":"Sentence"},{"id":"T780","span":{"begin":933,"end":1177},"obj":"Sentence"},{"id":"T781","span":{"begin":1178,"end":1287},"obj":"Sentence"},{"id":"T782","span":{"begin":1288,"end":1363},"obj":"Sentence"},{"id":"T783","span":{"begin":1364,"end":1514},"obj":"Sentence"},{"id":"T784","span":{"begin":1515,"end":1594},"obj":"Sentence"},{"id":"T785","span":{"begin":1595,"end":1658},"obj":"Sentence"},{"id":"T786","span":{"begin":1659,"end":1732},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Caco-2 cells seeded on glass coverslips were infected with SARS-CoV-2 Isolate Muc-IMB-1/2020, second passage on Vero E6 cells (2x106 PFU/mL) at an MOI of 0.1 or 0.01. At 24 hours post-infection cells were washed with PBS and fixed in 4% paraformaldehyde in PBS for 20 minutes at RT, followed by permeabilization with 0.3% Triton X-100 in PBS for 10 minutes at RT and blocking in 5% fetal calf serum in PBS for 1 hour at RT. Incubation with primary antibodies against CK2α (Abcam, ab70774, 1:500), SARS-CoV membrane (M) protein (Rockland, #100-401-A55, 1:500) and SARS-CoV nucleocapsid (N) protein (Rockland, #200-401-A50, 1:1000) was performed for 1 hour at RT. After washing with PBS, cells were incubated with AF568-labeled goat-anti-rabbit (Invitrogen, #A11011) and AF647-labeled goat-anti-mouse (Invitrogen, #A21235) secondary antibodies (1:200) as well as AF488-labeled Phalloidin (Hypermol, #8813-01, 1:250) for 2 hours at RT. Fluorescence images were generated using a LSM800 confocal laser-scanning microscope (Zeiss) equipped with a 63X, 1.4 NA oil objective and Airyscan detector and the Zen blue software (Zeiss) and processed with Zen blue software and ImageJ/Fiji. For 3D-reconstruction, cells were fixed and stained as indicated and imaged as z stack with 0.15 μm sections. Z stack was processed using Imaris software 64x 9.5.1 and displayed as MIP. To quantify colocalization between casein kinase II and N protein colocalization events in filopodia of SARS-CoV-2 infected Caco-2 cells were counted. 42 ± 19 % of the N protein particles detected in filopodia colocalize with CK2. Length of filopodia was measured using Metamorph (Version 7.8). Distance was measured starting at cortical actin to the tip of Filopodia."}