Fig. 7. NF-YA depletion activates a DNA damage signature in satellite cells.

a Enriched upregulated GO terms (>500 genes) related to cell death/apoptosis and DNA damage/integrity processes in NF-YA^cKO versus NF-YA^fl/fl SCs. Circles are colored by adjusted p value (degree of enrichment) and their size commensurate with the number of genes belonging to the relative term. b Heatmaps showing expression of MSigDB genes of DNA damage checkpoints among samples. High expressed genes are highlighted in green, whereas low expressed genes in red. c Relative mRNA levels measured by RT-qPCR analysis of the indicated genes in SCs isolated from NF-YA^cKO versus NF-YA^fl/fl mice. Data represent mean ± s.d. (two-tailed unpaired t-test: Mre11a p = 0.0172, Rad51 p = 0.0345, Brca1 p = 0.0274, Birc5 p = 0.0476, Bcl2l2 p = 0.0370; *p < 0.05, n = 5 mice). d Immunofluorescence (left panel) images and quantification of γH2AX + cells calculated as proportion on Pax7+ cells (right panel) in proliferating SCs cultures isolated from NF-YA^fl/fl and NF-YA^cKO. Data represent mean ± s.d. (two-tailed unpaired t-test: p = 0.0494; *p < 0.05, n = 6 mice). Scale bar: 100 µm. e Western blot analysis of DDR markers in whole-cell extracts of NF-YA^fl/fl and NF-YA^cKO isolated SCs. Total histone H2A and Vinculin were used as loading control. n = 3 experiments. f Relative transcript levels by RT-qPCR analysis of the Mre11 and Rad51 genes in NF-YA^fl/fl and NF-YA^cKO SCs transduced with empty or NF-YAl lentiviral particles. Data represent mean ± s.d. (one-way ANOVA: Mre11a F(2,9)=86.44 p < 0.0001; Rad51 F(2,9)=30.08 p = 0.0001. **p < 0.05, ***p < 0.01 vs NF-YA^fl/fl; ^##p < 0.05, ^###p < 0.001 vs empty). g Western blot analysis of DDR markers in NF-YA^cKO empty and NF-YAl-transduced satellite cells from two independent experiments (exp1 and exp2). Total histone H2A and Vinculin were used as loading control. n = 3 experiments.