Fig. 3: ZIKV-NS2A inhibits the production of CYP17A1 protein.

a, b HEK293T cells were co-transfected with plasmids encoding hCYP17A1-HA/mCYP17A1-HA and either ZIKV structural proteins C/E/M or nonstructural proteins NS1/2A/2B/3/4A/4B/5, fused with Flag or empty vector as control. The expression of hCYP17A1-HA/mCYP17A1-HA and individual ZIKV proteins was detected by Western blotting at 36 h post-transfection. The red font highlighted the ZIKV-NS2A protein. c, d HEK293T cells were co-transfected with plasmids encoding hCYP17A1-HA/mCYP17A1-HA and various amounts of NS2A-Flag. The expression of CYP17A1 and NS2A were detected by Western blotting at 36 h post-transfection. e CYP17A1-A549 cells were transfected with various amounts of NS2A-Flag. The expression of CYP17A1 and NS2A were detected by Western blotting at 36 h post-transfection. f CYP17A1-A549 cells were transfected with the NS2A-Flag or empty vector as control. At 36 h post-transfection, representative immunofluorescence images of these cells were shown by staining for NS2A-Flag and CYP17A1. DAPI staining indicates the nuclei. Scale bar, 10 μm. g, h Primary mLCs were transfected with various amounts of the NS2A-Flag. At 36 h post-transfection, the expression of endogenous CYP17A1/CYP11A1/STAR/3β-HSD (g) and CYP17A1 mRNA levels (h) were detected by Western blotting and qRT-PCR, respectively. The images in a–g are representative of three independent experiments. The data shown in h are the mean ± s.d (n = 4 independent experiments); Statistical analysis of (h) was performed using one-way ANOVA, followed by Tukey’s multiple comparison test; ns not significant. Source data are provided in the Source Data file.