Anti acetyl histone 3

ChIP was performed using the MAGnify Chromatin Immunoprecipitation System (Invitrogen). Protocol was performed as described in^{44 (link)}. For each ChIP reaction 300,000 cells were used, cell lysates were added to their respective antibody/beads for 2 h. Eluted DNA was PCR amplified with primers encompassing the Gli- responsive elements of human ABC promoter. The following antibodies were used: IgG rabbit (Invitrogen), rabbit polyclonal anti-Gli1 H300 (sc-20687, Santa Cruz Biotechnology Inc.), rabbit polyclonal anti-acetyl-histone 3 (06599, Millipore). Eluted DNA has been analysed with Q-PCR. Primers were designed with Primer-Blast designing tool (https://www.ncbi.nlm.nih.gov/tools/primer-blast/) and Primers tool (Genomatix Genome Analyzer, GGA, v3.30126, https://www.genomatix.de/) and are reported in Supplementary Table 2.

After 72 h of AA treatment cells were lysed by RIPA Lysis Buffer containing protease and phosphatase inhibitor. An equal amount of each protein sample was separated by 8–12% SDS–PAGE and transferred onto PVDF membranes. The following antibodies were incubated with the blocked membrane to detect the proteins of interest: anti-acetyl histone 3 (1:1000, Millipore 06-599), anti-histone 3 (1:2500, ABCAM ab1791), anti-PCAF (1:1000, Santa Cruz 13124) and β-actin (1:1000, Sigma-Aldrich A1978).