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. Author manuscript; available in PMC: 2017 Jun 6.
Published in final edited form as: Dev Cell. 2016 Jun 6;37(5):444–457. doi: 10.1016/j.devcel.2016.05.004

Figure4. FANCD2 promotes papillar cell viability and prevents micronucleus formation after IR.

Figure4

A.Representative image of papillar chromosomes in fancd2 RNAi#1 animal. Arrows indicate acentric chromosomes. DAPI indicates DNA. B.Frequency of acentric and fused chromosomes in WT vs. fancd2 RNAi#1 papillar cells +/− IR. C.Adult fancd2RNAi#1 rectum. D.Adult fancd2RNAi#1 rectum from animal irradiated during papillar endocycles. E.Adult fancd2RNAi#2 rectum from animal irradiated during papillar endocycles. Papillae false-colored in green, DNA in purple in C-E. For comparable controls see Fig3H,I. F.Adult papillar cell number in fancd2 +/−IR. N=8-30 animals/condition. Yellow bars=mean. *=significant change between NoIR vs. +IR compared to WT (Methods). G.Time-lapse of fancd2RNAi#1 papillar cell after IR. CenpC-Tomato=kinetochores (KTs, purple), histone H2AV=DNA (green, nuclear), and Moesin-GFP=cell membranes (Memb, green). Time is in min. relative to anaphase onset. White arrows=unaligned DNA that form micronuclei. Yellow arrow=cytokinetic furrow. H.Frequency unaligned DNA and micronuclei +/−IR in WT vs. fancd2RNAi#1. Data from 84-113 divisions/condition, from multiple replicates. *=significant change from no IR by Chi-squared (p<.00001, Methods). I.Unaligned PH3+ chromosomes (green labeling/arrows, DNA in purple) in fancd2RNAi#1 after IR. J.Persistent PH3+ micronuclei (green labeling/arrows, DNA in purple) in fancd2RNAi#1 after IR. K.Frequency PH3+ micronuclei +/−IR in WT vs. fancd2RNAi#1. From N=18-27 animals/condition from multiple replicates. Yellow bars=mean. *=significant change from no IR (Two-tailed T-test, p<.01). L.Diagram of acentric chromosome fate in fancd2 after IR. Green=DNA, Purple=Centromeres. Instead of segregating completely into daughter nuclei as in Fig3E, acentric DNA accumulates in micronuclei (indicated by small green circles without centromeres). DAPI=DNA in all images. White scale bar=50μm, yellow scale bar=5μm.