Fig. 2.

Mebendazole inhibits USP5 expression and induces c-Maf degradation via the ubiquitin-proteasome pathway. a LP1 and RPMI-8226 cells were treated with MBZ at the indicated concentrations for 24 h, followed by IB assays for c-Maf and USP5. Total RNA was extracted for RT-PCR to assess c-Maf and USP5 expression. GAPDH was used as the internal control. b The c-Maf and USP5 expression levels from a were analyzed by densitometry. c LP1 and RPMI-8226 cells were treated with MBZ or MG132 at the indicated concentrations for 24 h, followed by immunoblotting for c-Maf and GAPDH. d HEK293T cells were cotransfected with the HA-c-Maf and Myc-USP5 plasmids for 48 h, followed by MBZ treatment for 24 h and subsequent IP/IB assays. e LP1 and RPMI-8226 cells were treated with MBZ at the indicated concentrations for 24 h, followed by cell lysate preparation and IP/IB assays. f HEK293T cells were cotransfected with the HA-c-Maf, Myc-USP5 and Flag-Ub plasmids for 48 h. Cells were then treated with MBZ for 24 h before being lysed and subjected to IP/IB assays as indicated. g LP1 and RPMI-8226 cells were treated with MBZ at the indicated concentrations for 24 h, followed by IP/IB assays