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As reported before [2,3], Rb KO retinas exhibited both spatial and temporal ectopic DNA synthesis (Figures 1C and S1A). This is easily detected at E14, E16, and postnatal day (P) 0 in the inner retina where abnormal BrdU+ ganglion and amacrine RTCs are located, or on the outermost region of the P0 retina, where BrdU+ photoreceptor RTCs reside (Figures S1A and S2, arrows) [2]. Ectopic RTC division in Rb KO retinas is even more obvious at P8 or P18, when division is completed in wild-type (WT) retina (Figures 1C and S1A). Strikingly, the ectopically positioned S-phase cells at E14, E16, and P0 and all the abnormal division at P8 and P18 were completely suppressed in the Rb/E2f1 DKO retina (Figures 1C, 1E, 1F, S1A, and S2). In contrast, deletion of E2f2 or E2f3 had no effect at any stage of development. Analysis of mitotic cells with anti–phosphohistone 3 (PH3)–specific antibodies confirmed that loss of E2f1, but not E2f2 or E2f3, suppressed ectopic division (Figure S3). Deleting one E2f1 allele partially suppressed ectopic S-phase and mitosis in Rb KO RTCs (Figures 1C, 1E, 1F, S1A, S2, and S3), suggesting that E2f1 drives ectopic division in Rb KO RTCs in a dose-dependent fashion. These data contrast with previous findings in the lens and CNS of Rb KO embryos, where deletion of any activating E2f suppresses ectopic division to some extent [6–9]."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T6766","span":{"begin":19,"end":23},"obj":"CL_GO_EXT:cell"},{"id":"T6767","span":{"begin":19,"end":32},"obj":"GO:0051301"},{"id":"T6768","span":{"begin":34,"end":38},"obj":"NCBITaxon:10088"},{"id":"T6769","span":{"begin":50,"end":58},"obj":"CHEBI_SO_EXT:molecular_label_or_mark_or_tag_process"},{"id":"T6770","span":{"begin":64,"end":81},"obj":"CHEBI:472552"},{"id":"T6771","span":{"begin":83,"end":87},"obj":"CHEBI:472552"},{"id":"T6772","span":{"begin":100,"end":109},"obj":"GO_EXT:killing"},{"id":"T6773","span":{"begin":118,"end":135},"obj":"UBERON:0013682"},{"id":"T6774","span":{"begin":149,"end":153},"obj":"CHEBI:472552"},{"id":"T6775","span":{"begin":217,"end":219},"obj":"PR_EXT:000013773"},{"id":"T6776","span":{"begin":223,"end":230},"obj":"UBERON:0000966"},{"id":"T6777","span":{"begin":275,"end":278},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T6778","span":{"begin":275,"end":288},"obj":"GO:0071897"},{"id":"T6779","span":{"begin":356,"end":361},"obj":"GO:0007567"},{"id":"T6780","span":{"begin":385,"end":391},"obj":"UBERON:0000966"},{"id":"T6781","span":{"begin":407,"end":411},"obj":"CHEBI:472552"},{"id":"T6795","span":{"begin":871,"end":875},"obj":"PR_EXT:000006852"},{"id":"T6796","span":{"begin":880,"end":886},"obj":"UBERON:0000966"},{"id":"T6797","span":{"begin":935,"end":943},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T6798","span":{"begin":947,"end":951},"obj":"PR_EXT:000006853"},{"id":"T6799","span":{"begin":955,"end":959},"obj":"PR_EXT:000006854"},{"id":"T6800","span":{"begin":1015,"end":1022},"obj":"GO:0007067"},{"id":"T6801","span":{"begin":1023,"end":1028},"obj":"CL_GO_EXT:cell"},{"id":"T6802","span":{"begin":1039,"end":1053},"obj":"CHEBI_EXT:phosphohistone"},{"id":"T6803","span":{"begin":1071,"end":1081},"obj":"GO:0042571"},{"id":"T6804","span":{"begin":1105,"end":1109},"obj":"PR_EXT:000006852"},{"id":"T6805","span":{"begin":1119,"end":1123},"obj":"PR_EXT:000006853"},{"id":"T6806","span":{"begin":1127,"end":1131},"obj":"PR_EXT:000006854"},{"id":"T6807","span":{"begin":1174,"end":1182},"obj":"SO_EXT:sequence_deletion_process"},{"id":"T6808","span":{"begin":1187,"end":1191},"obj":"PR_EXT:000006852"},{"id":"T6809","span":{"begin":1192,"end":1198},"obj":"SO_EXT:0001023"},{"id":"T6810","span":{"begin":1228,"end":1235},"obj":"GO:0051320"},{"id":"T6811","span":{"begin":1240,"end":1247},"obj":"GO:0007067"},{"id":"T6812","span":{"begin":1251,"end":1253},"obj":"PR_EXT:000013773"},{"id":"T6813","span":{"begin":1317,"end":1321},"obj":"PR_EXT:000006852"},{"id":"T6814","span":{"begin":1349,"end":1351},"obj":"PR_EXT:000013773"},{"id":"T6815","span":{"begin":1439,"end":1443},"obj":"UBERON_EXT:lens"},{"id":"T6816","span":{"begin":1448,"end":1451},"obj":"UBERON:0001017"},{"id":"T6817","span":{"begin":1455,"end":1457},"obj":"PR_EXT:000013773"},{"id":"T6818","span":{"begin":1461,"end":1468},"obj":"UBERON:0000922"},{"id":"T6819","span":{"begin":1476,"end":1484},"obj":"SO_EXT:sequence_deletion_entity_or_process"},{"id":"T6820","span":{"begin":1503,"end":1506},"obj":"PR_EXT:E2F"},{"id":"T6782","span":{"begin":413,"end":421},"obj":"UBERON:0000045"},{"id":"T6783","span":{"begin":426,"end":434},"obj":"CL:0000561"},{"id":"T6784","span":{"begin":490,"end":496},"obj":"UBERON:0000966"},{"id":"T6785","span":{"begin":504,"end":508},"obj":"CHEBI:472552"},{"id":"T6786","span":{"begin":510,"end":523},"obj":"CL:0000210"},{"id":"T6787","span":{"begin":594,"end":596},"obj":"PR_EXT:000013773"},{"id":"T6788","span":{"begin":600,"end":607},"obj":"UBERON:0000966"},{"id":"T6789","span":{"begin":673,"end":682},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T6790","span":{"begin":684,"end":686},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T6791","span":{"begin":688,"end":694},"obj":"UBERON:0000966"},{"id":"T6792","span":{"begin":756,"end":763},"obj":"GO:0051320"},{"id":"T6793","span":{"begin":764,"end":769},"obj":"CL_GO_EXT:cell"},{"id":"T6794","span":{"begin":868,"end":870},"obj":"PR_EXT:000013773"}],"text":"To measure ectopic cell division, mice were pulse-labelled with bromodeoxyuridine (BrdU) 2 h before sacrifice and the peripheral retina analyzed for BrdU incorporation by immunofluorescence. As reported before [2,3], Rb KO retinas exhibited both spatial and temporal ectopic DNA synthesis (Figures 1C and S1A). This is easily detected at E14, E16, and postnatal day (P) 0 in the inner retina where abnormal BrdU+ ganglion and amacrine RTCs are located, or on the outermost region of the P0 retina, where BrdU+ photoreceptor RTCs reside (Figures S1A and S2, arrows) [2]. Ectopic RTC division in Rb KO retinas is even more obvious at P8 or P18, when division is completed in wild-type (WT) retina (Figures 1C and S1A). Strikingly, the ectopically positioned S-phase cells at E14, E16, and P0 and all the abnormal division at P8 and P18 were completely suppressed in the Rb/E2f1 DKO retina (Figures 1C, 1E, 1F, S1A, and S2). In contrast, deletion of E2f2 or E2f3 had no effect at any stage of development. Analysis of mitotic cells with anti–phosphohistone 3 (PH3)–specific antibodies confirmed that loss of E2f1, but not E2f2 or E2f3, suppressed ectopic division (Figure S3). Deleting one E2f1 allele partially suppressed ectopic S-phase and mitosis in Rb KO RTCs (Figures 1C, 1E, 1F, S1A, S2, and S3), suggesting that E2f1 drives ectopic division in Rb KO RTCs in a dose-dependent fashion. These data contrast with previous findings in the lens and CNS of Rb KO embryos, where deletion of any activating E2f suppresses ectopic division to some extent [6–9]."}

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{"project":"2_test","denotations":[{"id":"17608565-15193257-84868971","span":{"begin":211,"end":212},"obj":"15193257"},{"id":"17608565-15231717-84868972","span":{"begin":213,"end":214},"obj":"15231717"},{"id":"17608565-15193257-84868973","span":{"begin":566,"end":567},"obj":"15193257"},{"id":"17608565-9774968-84868974","span":{"begin":1551,"end":1552},"obj":"9774968"},{"id":"17608565-11146559-84868974","span":{"begin":1551,"end":1552},"obj":"11146559"},{"id":"17608565-11230146-84868974","span":{"begin":1551,"end":1552},"obj":"11230146"},{"id":"17608565-12065245-84868974","span":{"begin":1551,"end":1552},"obj":"12065245"},{"id":"T7044","span":{"begin":211,"end":212},"obj":"15193257"},{"id":"T88570","span":{"begin":213,"end":214},"obj":"15231717"},{"id":"T61973","span":{"begin":566,"end":567},"obj":"15193257"},{"id":"T60221","span":{"begin":1551,"end":1552},"obj":"9774968"},{"id":"T11659","span":{"begin":1551,"end":1552},"obj":"11146559"},{"id":"T49571","span":{"begin":1551,"end":1552},"obj":"11230146"},{"id":"T50861","span":{"begin":1551,"end":1552},"obj":"12065245"}],"text":"To measure ectopic cell division, mice were pulse-labelled with bromodeoxyuridine (BrdU) 2 h before sacrifice and the peripheral retina analyzed for BrdU incorporation by immunofluorescence. As reported before [2,3], Rb KO retinas exhibited both spatial and temporal ectopic DNA synthesis (Figures 1C and S1A). This is easily detected at E14, E16, and postnatal day (P) 0 in the inner retina where abnormal BrdU+ ganglion and amacrine RTCs are located, or on the outermost region of the P0 retina, where BrdU+ photoreceptor RTCs reside (Figures S1A and S2, arrows) [2]. Ectopic RTC division in Rb KO retinas is even more obvious at P8 or P18, when division is completed in wild-type (WT) retina (Figures 1C and S1A). Strikingly, the ectopically positioned S-phase cells at E14, E16, and P0 and all the abnormal division at P8 and P18 were completely suppressed in the Rb/E2f1 DKO retina (Figures 1C, 1E, 1F, S1A, and S2). In contrast, deletion of E2f2 or E2f3 had no effect at any stage of development. Analysis of mitotic cells with anti–phosphohistone 3 (PH3)–specific antibodies confirmed that loss of E2f1, but not E2f2 or E2f3, suppressed ectopic division (Figure S3). Deleting one E2f1 allele partially suppressed ectopic S-phase and mitosis in Rb KO RTCs (Figures 1C, 1E, 1F, S1A, S2, and S3), suggesting that E2f1 drives ectopic division in Rb KO RTCs in a dose-dependent fashion. These data contrast with previous findings in the lens and CNS of Rb KO embryos, where deletion of any activating E2f suppresses ectopic division to some extent [6–9]."}