Western Blot Analysis of Signaling Proteins

Western blot analysis was carried out as previously described [30 (link)]. The following primary antibodies (Abs) were used: anti-phospho-H2A.X (S139) (1/2000), anti-H2A.X (1/1000), anti-phospho-ERK1/2 (Thr202/Tyr204) (1/2000), anti-ERK1/2 (1/2000), anti-phospho-MEK1/2 (Ser217/221) (1/2000), and anti-MEK1/2 (1/2000) from Cell Signaling Technology (Danvers, MA, USA), and anti-PARP-1 (1/200; Santa Cruz Biotechnology, (Dallas, TX, USA)) and anti-Actin (1/10,000; Sigma-Aldrich). The following secondary Abs were used: anti-rabbit-HRP (1/5000) and anti-mouse-HRP (1/5000 and 1/10,000 for actin) from SouthernBiotech (Nanterre, France). The Western blots were scanned using ChemiDoc imaging system (Bio-Rad, Marnes-la-Coquette, France). The densitometric analyses were performed using an ImageJ software (National Institutes of Health, Bethesda, MD, USA) (accessed on 11 November 2020).