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c accumulation in homozygous CbCln3Δex7/8 cerebellar precursor cells a. Subunit c immunostaining and autofluorescence of 7-day confluency-aged wild-type and homozygous CbCln3Δex7/8 cells is shown. Wild-type cultures (CbCln3+/+) exhibited limited subunit c immunostaining and autofluorescence. However, CbCln3Δex7/8/Δex7/8 cells contained numerous subunit c puncta. Autofluorescence (7 days AF) was also significantly elevated (right panels), although limited overlap with subunit c puncta was observed (arrows). 40 × magnification. b. Immunoblot analysis of subunit c protein at sub-confluency or 7-day confluency incubation is shown. Total protein extracts from sub-confluency wild-type (+/+) and homozygous mutant (Δex7/8/Δex7/8) cultures contained approximately equal levels of subunit c protein (α-sub c). 7-day confluency extract from homozygous CbCln3Δex7/8 cells (Δex7/8/Δex7/8) had elevated levels of subunit c protein (~1.5X), relative to wild-type extract (+/+). Protein levels were normalized to cytochrome c oxidase subunit IV (α-cox4). c. TEM analysis of inclusions in 7-day confluency-aged homozygous CbCln3Δex7/8 cells is shown. A large autophagosome contained by double membrane (arrows) is filled with degenerating mitochondria (Md), electron dense cores (left and right of *) and other smaller vesicular structures. A large electron-dense inclusion, with a lipofuscin (Ln) appearance, is also present. M, mitochondria. 10,000 × magnification."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T8745","span":{"begin":0,"end":9},"obj":"PR_EXT:000022190"},{"id":"T8746","span":{"begin":37,"end":39},"obj":"UBERON:0002037"},{"id":"T8747","span":{"begin":39,"end":43},"obj":"PR_EXT:000005591"},{"id":"T8748","span":{"begin":50,"end":60},"obj":"UBERON:0002037"},{"id":"T8749","span":{"begin":71,"end":76},"obj":"CL_GO_EXT:cell"},{"id":"T8750","span":{"begin":80,"end":89},"obj":"PR_EXT:000022190"},{"id":"T8751","span":{"begin":146,"end":150},"obj":"GO_PATO_EXT:aging_or_agedness"},{"id":"T8752","span":{"begin":151,"end":160},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T8753","span":{"begin":176,"end":178},"obj":"UBERON:0002037"},{"id":"T8754","span":{"begin":178,"end":182},"obj":"PR_EXT:000005591"},{"id":"T8755","span":{"begin":189,"end":194},"obj":"CL_GO_EXT:cell"},{"id":"T8756","span":{"begin":205,"end":214},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T8757","span":{"begin":225,"end":227},"obj":"UBERON:0002037"},{"id":"T8758","span":{"begin":227,"end":231},"obj":"PR_EXT:000005591"},{"id":"T8759","span":{"begin":231,"end":232},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T8760","span":{"begin":233,"end":234},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T8761","span":{"begin":254,"end":263},"obj":"PR_EXT:000022190"},{"id":"T8762","span":{"begin":310,"end":312},"obj":"UBERON:0002037"},{"id":"T8763","span":{"begin":312,"end":316},"obj":"PR_EXT:000005591"},{"id":"T8764","span":{"begin":330,"end":335},"obj":"CL_GO_EXT:cell"},{"id":"T8765","span":{"begin":355,"end":364},"obj":"PR_EXT:000022190"},{"id":"T8766","span":{"begin":480,"end":489},"obj":"PR_EXT:000022190"},{"id":"T8767","span":{"begin":566,"end":575},"obj":"PR_EXT:000022190"},{"id":"T8768","span":{"begin":576,"end":583},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8769","span":{"begin":649,"end":656},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8770","span":{"begin":686,"end":695},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T8771","span":{"begin":697,"end":698},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T8772","span":{"begin":699,"end":700},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T8773","span":{"begin":717,"end":723},"obj":"SO_EXT:sequence_altered_entity_or_alteration_process"},{"id":"T8774","span":{"begin":789,"end":798},"obj":"PR_EXT:000022190"},{"id":"T8775","span":{"begin":799,"end":806},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8776","span":{"begin":810,"end":815},"obj":"PR_EXT:000022190"},{"id":"T8777","span":{"begin":859,"end":861},"obj":"UBERON:0002037"},{"id":"T8778","span":{"begin":861,"end":865},"obj":"PR_EXT:000005591"},{"id":"T8779","span":{"begin":872,"end":877},"obj":"CL_GO_EXT:cell"},{"id":"T8780","span":{"begin":917,"end":926},"obj":"PR_EXT:000022190"},{"id":"T8781","span":{"begin":927,"end":934},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8782","span":{"begin":956,"end":965},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T8783","span":{"begin":975,"end":976},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T8784","span":{"begin":977,"end":978},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T8785","span":{"begin":981,"end":988},"obj":"CHEBI_PR_EXT:protein"},{"id":"T8786","span":{"begin":1015,"end":1027},"obj":"CHEBI:18070"},{"id":"T8787","span":{"begin":1015,"end":1035},"obj":"GO_EXT:0004129"},{"id":"T8788","span":{"begin":1015,"end":1046},"obj":"PR_EXT:000005776"},{"id":"T8789","span":{"begin":1050,"end":1054},"obj":"PR_EXT:000005776"},{"id":"T8790","span":{"begin":1076,"end":1086},"obj":"GO:0016234"},{"id":"T8791","span":{"begin":1107,"end":1111},"obj":"GO_PATO_EXT:aging_or_agedness"},{"id":"T8792","span":{"begin":1123,"end":1125},"obj":"UBERON:0002037"},{"id":"T8793","span":{"begin":1125,"end":1129},"obj":"PR_EXT:000005591"},{"id":"T8794","span":{"begin":1136,"end":1141},"obj":"CL_GO_EXT:cell"},{"id":"T8795","span":{"begin":1160,"end":1173},"obj":"GO:0005776"},{"id":"T8796","span":{"begin":1194,"end":1202},"obj":"GO:0016020"},{"id":"T8797","span":{"begin":1240,"end":1252},"obj":"GO:0005739"},{"id":"T8798","span":{"begin":1259,"end":1267},"obj":"CHEBI:10545"},{"id":"T8799","span":{"begin":1259,"end":1279},"obj":"GO:0031045"},{"id":"T8800","span":{"begin":1320,"end":1329},"obj":"GO:0031982"},{"id":"T8801","span":{"begin":1350,"end":1358},"obj":"CHEBI:10545"},{"id":"T8802","span":{"begin":1365,"end":1374},"obj":"GO:0016234"},{"id":"T8803","span":{"begin":1428,"end":1429},"obj":"GO:0005739"},{"id":"T8804","span":{"begin":1431,"end":1443},"obj":"GO:0005739"}],"text":"Subunit c accumulation in homozygous CbCln3Δex7/8 cerebellar precursor cells a. Subunit c immunostaining and autofluorescence of 7-day confluency-aged wild-type and homozygous CbCln3Δex7/8 cells is shown. Wild-type cultures (CbCln3+/+) exhibited limited subunit c immunostaining and autofluorescence. However, CbCln3Δex7/8/Δex7/8 cells contained numerous subunit c puncta. Autofluorescence (7 days AF) was also significantly elevated (right panels), although limited overlap with subunit c puncta was observed (arrows). 40 × magnification. b. Immunoblot analysis of subunit c protein at sub-confluency or 7-day confluency incubation is shown. Total protein extracts from sub-confluency wild-type (+/+) and homozygous mutant (Δex7/8/Δex7/8) cultures contained approximately equal levels of subunit c protein (α-sub c). 7-day confluency extract from homozygous CbCln3Δex7/8 cells (Δex7/8/Δex7/8) had elevated levels of subunit c protein (~1.5X), relative to wild-type extract (+/+). Protein levels were normalized to cytochrome c oxidase subunit IV (α-cox4). c. TEM analysis of inclusions in 7-day confluency-aged homozygous CbCln3Δex7/8 cells is shown. A large autophagosome contained by double membrane (arrows) is filled with degenerating mitochondria (Md), electron dense cores (left and right of *) and other smaller vesicular structures. A large electron-dense inclusion, with a lipofuscin (Ln) appearance, is also present. M, mitochondria. 10,000 × magnification."}