Fig. 4: HDAC3 depletion in FP-RMS cell lines combined to IR induces DNA damage and impairs DNA repair. | Cell Death Discovery

Fig. 4: HDAC3 depletion in FP-RMS cell lines combined to IR induces DNA damage and impairs DNA repair.

From: HDAC3 genetic and pharmacologic inhibition radiosensitizes fusion positive rhabdomyosarcoma by promoting DNA double-strand breaks

Fig. 4

A Representative immunofluorescence images of γH2AX foci (green, left) (n = 3 independent biological replicates) in HDAC3 depleted -RH30 and -RH4 cells 6 h post IR with 6 Gy. DAPI (blue) was used as nuclear counterstain. Images were taken using confocal microscopy with a ×60 oil immersion objective lens. Scale Bar = 10 μm. Scatter plot of γH2AX average intensity per cell number (right) in HDAC3 depleted -RH30 and -RH4 cells after 6 h post IR with 6 Gy. Graph represents the mean of three independent experiments ±SEM, ANOVA one-way test. Exact p-values are reported in the figure. B Analysis of DSBs accumulation by the neutral Comet assay. Representative images are reported above. RH30 and RH4 HDAC3 KO cells were exposed or not to 6 Gy of IR and after 6 h subjected to the neutral Comet assay. In the scatter plot below, data are presented as mean tail moment ±SEM, ANOVA one-way test. Exact p values are reported in the figure. C Representative western blot (n = 3 independent biological replicates) depicting the effect of IR (6 Gy), HDAC3 depletion and their combination on DNA damage. The cells were processed 6 h post IR. pATM (Ser 1981), total ATM, pDNA-PKcs (Thr2609), total pDNA-PKcs, Rad51, and Ku70 protein levels were detected. All protein levels were normalized on Vinculin.

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