Fig. 7
From: Non-canonical reader modules of BAZ1A promote recovery from DNA damage
Function-altering PHD substitutions are detrimental to the cellular functions of BAZ1A. a Western blot analysis of BAZ1A expression in parental, BAZ1A-KO, and BAZ1A-KO cells engineered to re-express BAZ1AKAKY. b–d Confluence of the indicated cell line measured over time, before, and after a pulse of UVC light. c Accumulation of endogenous SMARCA5 at sites of DNA damage as in Fig. 2c, d. Scale bar, 20 μm. Histograms represent the mean value ± s.e.m. normalized to recruitment in parental cells. P-values were calculated using a two-tailed Mann–Whitney test; n.s. not significant (P > 0.05). d Accumulation of GFP-fusion BAZ1A proteins to sites of laser irradiation in live cells. The fluorescence signal at site of irradiation is normalized to the signal of a non-irradiated area. Signal above 1 indicates accumulation at site of damage. Numbers on the x-axis and y-axis represent the time at which the fluorescence value equals 1 and the maximum value (plateau determined by fitting), respectively. e PC analysis of the transcriptional profiles of parental, BAZ1A-KO, re-expressing BAZ1A wild-type and re-expressing BAZ1AKAKY cells, before and after DNA damage. For each cell line, the first and second PCs (explaining the highest percent of variance) are reported for 2–3 independent RNA extractions. f Heat map showing the expression profiles of genes that are significantly upregulated in parental and re-expressing BAZ1A cells after UVC treatment compared to BAZ1AKAKY and BAZ1A-KO cells. The genes that are significantly downregulated for the same comparison are shown in Supplementary Fig. 8d. In both cases, only genes with the same expression profiles in parental and re-expressing BAZ1A cells, as well as BAZ1A-KO and BAZ1AKAKY cells are reported. Individual columns correspond to independent RNA extractions. g 4-way comparison showing the log2 fold-change between BAZ1A-KO cells engineered to re-express BAZ1A wild-type or the BAZ1AKAKY mutant under control conditions on the x-axis compared to UVC conditions on the y-axis. Points are colored according to differential expression significance, points in gray do not satisfy significance cut-off (P < 0.01, absolute log2 fold-change >1)
