Ep1713y

Protein isolation and western blots were performed as described before [48 (link)]. For analysis of phosphorylated proteins, cells were harvested and lysed with radioimmunoprecipitation assay (RIPA) buffer (89900, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with protease and phosphatase inhibitors (cOmpleteTM Tablets, 11697498001, Roche, Basel, Swiss; Sodium orthovanadate, 13721-39-6, Fivephoton Biochemicals, San Diego, CA, USA). Antibodies against rabbit anti-GLI1 monoclonal antibody (C68H3, 1:1000, Cell Signaling Technology, Inc., Danvers, MA, USA), mouse anti-GLI-2 (Dilution, C-10: sc-271786, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit Anti-HSF1-phospho(S326) antibody (1:5000, EP1713Y, abcam, Cambridge, UK), rabbit Anti-HSF1 antibody (1:1000, 4356, Cell Signaling Technology, Inc., Danvers, MA, USA), Mouse anti-HSP70/HSPA1A antibody (1:1000, MAB1663, R&D Systems, Inc. Minneapolis, MN, USA), Rabbit anti-HSP90 Antibody (1:1000, #4874, Cell Signaling Technology, Inc., Danvers, MA, USA) or mouse anti-β-ACTIN (sc-47778, 1:5000, Santa Cruz Biotechnology, Dallas, TX, USA). Secondary antibodies against HRP-linked anti-rabbit immunoglobulins (1:10,000, 7074) and anti-mouse IgG (1:10,000, NXA931) secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) and GE Healthcare (Cytiva, Marlborough, MA, USA), respectively.