Anti p h2ax

The tumors were fixed for 36 h in 4% paraformaldehyde, transferred to 70% EtOH and paraffin-embedded. Sections 4 µm thick were deparaffinized, rehydrated and incubated for 20 min at 37 °C with anti-P-EGFR (1–100, ab40815, Abcam, Cambridge, UK) or anti-P-H2AX (1–100, sc-517348, Santa Cruz Biotechnology, Dallas, TX, USA), followed by secondary Ab incubation with OmniMap anti-Rb HRP (760–4311, Roche, Basel, Switzerland) at room temperature for 16 min, followed by the detection kit ChromoMap Kit (760–4304, Roche, Basel, Switzerland), and the slides were then counterstained with hematoxylin, dehydrated, cleared and cover-slipped. The slides were digitally scanned using an Aperio scanner scope XT.

The following antibodies were used: anti-Ki67 (ab16667, Abcam; IF), anti-BrdU (347580, Biosciences; IF), anti-filaggrin (PRB-417, Covance; IF), anti-involucrin (RINVOL, 924401, Biologend; IF), anti-K5 (SAB45016501, Sigma-Aldrich; IF), anti-K10 (sc-23877, Santa Cruz Biotechnology; IF), anti-K16 (sc-53255, Santa Cruz Biotechnology; IF), anti-Cyclin A (sc-751, Santa Cruz Biotechnology; IF), anti-pH3 (sc-8656-R, Santa Cruz Biotechnology; IF), anti-p-H2AX (sc-517348, Santa Cruz Biotechnology; IF), anti-p40 (deltaNp63, 24-8626-RBP1, ARP; IF), anti-Cyclin E1 (sc-248, Santa Cruz Biotechnology; WB), anti-PCNA (sc-56, Santa Cruz Biotechnology; WB) and anti-GAPDH (sc-32233, Santa Cruz Biotechnology; WB).The following secondary antibodies were used: Alexa Fluor® 488-conjugated goat anti-rabbit or anti-mouse IgG antibodies (Jackson; IF), Alexa Fluor® 594-conjugated goat anti-rabbit or anti-mouse IgG antibodies (Jackson; IF) and DyLight^TM 800-conjugated goat anti-mouse IgG antibody (Invitrogen; WB).