Fig. 4

TRPM1 associated with the HSP90 chaperone complex. a Representative western blot results of 3xF-TRPM1, TRPM1, HSP70 and CDC37 co-immunoprecipitated (co-IP) with HSP90. Cell lysates from Mel1617 cells stably expressing either an empty vector or 3xF-TRPM1 were immunoprecipitated (IP) with anti-HSP90 conjugated agarose. Mouse IgG antibodies were served as an IP control. n = 3. b Representative western blot results of HSP90 and HSP70 co-IP with 3xF-TRPM1. Mel1617 cells stably expressing 3xF-TRPM1 were mock-treated (−) or treated with 30 nM AUY922 for 6 h (+), cell lysates were prepared and IP with anti-FLAG M2 affinity gel. Cells expressing an empty vector were served as a control. n = 3. c Representative western blot results of HSP90 and HSP70 co-IP with TRPM1. Mel1617 cells were mock-treated (−) or treated with 30 nM AUY922 for 6 h (+), cell lysates were prepared and IP with anti-TRPM1 antibodies. Mouse IgG antibodies were served as an IP control. n = 3. d Representative western blot results of CDC37, TRPM1 and HSP70 co-IP with HSP90. Mel1617 cells were mock-treated or treated with 30 nM AUY922 for 6 h (+), cell lysates were prepared and IP with anti-HSP90 conjugated agarose. Mouse IgG antibodies were served as an IP control. n = 3. e Representative western blot results of Mel1617 cells expressing either a scrambled shRNA or shRNAs specific for CDC37. β-Actin was used as a loading control. n = 3