Figure 3. Suppressing ORP2 alters cellular steroid metabolite concentrations.

H295R WT and ORP2^kd cells were cultured into 100 mm dishes treated for 48 h with 0.4 mM Bt₂cAMP media collected and the amounts of steroids quantified by ultra performance liquid chromatography tandem mass spectrometry as described in Materials and Methods. Data are displayed as a heat map of the steroid hormone biosynthetic pathway, with steroid metabolites in untreated cells shown in A and Bt₂cAMP metabolites depicted in B. Fold change in metabolite secretion ORP2^kd is normalized to the wild type and data represent the mean ± SD of two separate experiments (n=6 per experiment). Steroid metabolite amounts are normalized to the cellular protein concentration. Enzymes catalyzing each reaction are denoted adjacent to the arrows. Abbreviations are as follows: cholesterol, CHOL; P5, pregnenolone; 17-OHP5, 17-hydroxypregnenolone; DHEA, dehydroepiandrosterone; P4, progesterone; 17-OHP4, 17-hydroxyprogesterone; A4, androstenedione; T, testosterone; DCORT, deoxycorticosterone; DCRT, 11-deoxycortisol; E1, estrone; E2, estradiol; CORT, corticosterone; S, 11-deoxycortisol; F, cortisol; ALDO, aldosterone.