PMC:5985359 / 11050-12279 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/5985359","sourcedb":"PMC","sourceid":"5985359","source_url":"https://www.ncbi.nlm.nih.gov/pmc/5985359","text":"Immunocytochemistry (ICC)\nCECs grown on glass coverslips were fixed with 4% paraformaldehyde (PFA) in PBS for 10 min. After washing with PBS, cells were permeabilized with 0.1% Triton X-100 for 10 min and non-specific binding sites were blocked in PBS with 3% BSA and 10% donkey serum for 1 hr. The CECs were incubated with primary antibodies, diluted to the appropriate concentration (Table S1), in blocking solution overnight at 4°C. For MBNL127 and MBNL2 antibodies, 0.5% Triton X-100 was incorporated in the blocking solution instead, omitting the previous permeabilization step, and proceeding normally after. After washing with PBS, CECs were incubated in donkey fluorophore-bound secondary antibodies (Alexa Fluor 488, anti-mouse or anti-rabbit; Invitrogen) diluted 1:1,000 in 3% BSA PBS for 1 hr at room temperature. Cells were washed again and incubated in DAPI stain (1:5,000 dilution; Sigma) for 2 min. Finally, coverslips were mounted onto microscope slides using Fluorescent Mounting Medium (Dako). Appropriate negative controls were carried out by performing the same protocol without the addition of primary antibodies. Images were taken using a confocal Zeiss 700 microscope and processed with the Zeiss software.","divisions":[{"label":"title","span":{"begin":0,"end":25}}],"tracks":[{"project":"2_test","denotations":[{"id":"29526280-16717059-2044774","span":{"begin":444,"end":447},"obj":"16717059"}],"attributes":[{"subj":"29526280-16717059-2044774","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#93e7ec","default":true}]}]}}