Fig. 5.

Cmpd 81 induces HMGCR ubiquitination and degradation. a, b CHO-7 cells were treated with Cmpd 81 at indicated concentrations for 16 h. 24,25-DHL was used as a positive control. a Immunoblotting analysis of HMGCR protein. b Quantification of Cmpd 81-induced HMGCR degradation shown in a. HMGCR protein of DMSO-treated cells was defined as 100. The EC₅₀ value of Cmpd 81 was 0.39 μM. c CHO-7 cells were treated with Cmpd 81 in the presence (+) or absence (−) of 10 μM MG-132 for 5 h. MG-132 prevented Cmpd 81-induced HMGCR degradation. d CHO-7 cells were treated with Cmpd 81 and 10 μM MG-132 for 3 h. Lysates were immunoprecipitated and pellets were probed for anti-ubiquitin (P4D1) and anti-HMGCR (A9). e CHO-7 cells were transfected with indicated plasmids and treated as in d. Lysates were immunoprecipitated with anti-Flag M2 beads, and pellets were probed for anti-ubiquitin (HA) and anti-HMGCR (Flag). f CHO-7 cells were transfected with indicated plasmids, then treated with Cmpd 81 for 5 h. Cmpd 81 promoted the degradation of WT but not ubiquitin sites mutated (K89R/K248R) HMGCR protein. g CHO-7 and SRD15 cells were treated with Cmpd 81 and the HMGCR protein was subjected for immunoblotting analysis. Data are from three independent experiments and presented as mean ± SD. Source data are provided as a Source Data File. Uncropped immunoblots are shown in Supplementary Fig. 10