Fig. 2

The C987X residue is critical for CYP17A1 stability. A HEK 293T cells were transfected with plasmids encoding CYP17A1(WT) or CYP17A1(C987X). After 48 hours, cells were treated with 100 μmol/L cycloheximide (CHX) for the indicated periods. B Densitometric analysis of CYP17A1(WT) and CYP17A1(C987X) proteins shown in (A). The densitometry of CYP17A1(WT) protein at 0 hour is defined as 1. C HEK293T cells were transfected with indicated plasmids and treated with 10 μm MG132 for 30 minutes. Cells were harvested, CYP17A1(WT) or CYP17A1(C987X) was immunoprecipitated by the anti-Myc coupled agarose and probed for the anti-HA antibody. Results shown are representative of two independent experiments. D Alignment of sequences flanking the C987X residue of CYP17A1 in various species. E Subcellular localization of CYP17A1(WT) or CYP17A1(C987X). HEK 293T cells were transfected with a plasmid encoding Myc-FLAG and stained with anti-FLAG and anti-KDEL antibodies. Scale bar =10 μm. F HEK293T cells were transfected with a plasmid encoding CYP17A1-FLAG and stained with anti-Myc tag and anti-COX IV antibodies, followed by microscopic analysis of CYP17A1 expression using a confocal microscope