Figure 5.

Levels of Endogenous CYP51A1 and TMUB2 Are Regulated by the RNF185-MBRL Complex

(A and B) Protein extracts from RNF185, MBRL, and HRD1 KO (A) HEK293 and (B) HeLa cells expressing either an EV or cDNAs encoding the WT or CI version of the indicated protein were analyzed by SDS-PAGE and immunoblotting. Extracts from parental cells left untreated or treated with the p97 inhibitor CB-5083 (CB) (4 h; 2.5 μM) were also analyzed. Relative CYP51A1 and TMUB2 levels are displayed below the respective blot, normalized to the GAPDH loading control.

(C) Endogenous CYP51A1 co-precipitates specifically with RNF185/MBRL complex. The indicated FLAG-tagged proteins (x axis) were precipitated from untreated or CB-5083-treated cells, and eluted proteins were analyzed by mass spectrometry. The y axis shows the log2 fold enrichment of endogenous CYP51A1 in the FLAG precipitates versus untagged control.

See also Figure S5.