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. 2015 Nov 26;6:8966. doi: 10.1038/ncomms9966

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(a,c) Western blots show significantly increased H2Av-pSer137 and p53 levels in GFAP^R79H transgenic flies. The blots were reprobed for actin to illustrate equivalent loading. *P=0.0290, t=3.034, df=5, n=6 (a); *P=0.0356, t=3.118, df=4, n=5 (c); unpaired t-test. Ctrl: repo-GAL4/+. GFAP^R79H: repo-GAL4, UAS-GFAP^R79H/+. (b,d) H2Av-pS137 and p53 colocalize with the glial cell marker Repo (top panels, arrows) but not with the neuronal cell marker elav (bottom panels, arrowheads) in GFAP^R79H transgenic flies. 4,6-Diamidino-2-phenylindole (DAPI) shows nuclei. Scale bar, 5 μm. Genotype: repo-GAL4, UAS-GFAP^R79H/+. (e) Double-label immunofluorescence reveals colocalization of H2Av-pS137 and p53 in GFAP^R79H transgenic flies (arrows). DAPI labels nuclei. Scale bar, 5 μm. Genotype: repo-GAL4, UAS-GFAP^R79H/+. (f) Knocking down or removing p53 with transgenic RNAi or a p53^null allele decreases the number of TUNEL-positive cells in GFAP^R79H transgenic flies. **P<0.0001, F=74.69, df=18; one-way ANOVA, n⩾6 per genotype. Genotypes are indicated in the figure label. Driver: repo-GAL4. (g) Seizure frequency is decreased with reduced expression of p53 (p53 RNAi). **P<0.0001, χ²=45.671, df=1; χ²-test, n>100 per genotype. Genotypes are indicated in the figure label. Driver: repo-GAL4. (h) Overexpression of Nos (GFAP Nos OE) increases, while reducing Nos (GFAP Nos RNAi) decreases the number of p53-positive cells in GFAP transgenic flies. No p53 accumulation is present in Ctrl and Nos OE alone. *P<0.01, **P<0.0001, F=69.03, df=30; one-way ANOVA, n⩾6 per genotype. Genotypes are indicated in the figure label. Driver: repo-GAL4. Ctrl: repo-GAL4/+. Flies were 1 day old in g and 20 days old in other panels.