FIG. 2.

Effects of in utero exposure to genistein (GEN) and DEHP on steroidogenic mediators. Relative mRNA expression of steroidogenic enzymes Cyp11a1 (A) and Hsd3b (B), and androgen receptor (Ar) (C) in PND3 and PND6 testes. Data are expressed as mean relative mRNA levels ± SEM normalized to alpha-tubulin (n = 4 per treatment). To further validate mRNA alterations, immunohistochemical analysis of HSD3B (D) was performed on PND3 testes and positive interstitial staining was quantified by image analysis (E); photos taken at 40× magnification. Immunohistochemical quantification is represented by mean fold change ± SEM of HSD3B staining relative to control (n ≥ 3). Omission of the primary antibody was used as a negative control (data not shown). Representative pictures are presented. Asterisks in both mRNA and protein analyses indicate a significant difference relative to control (P ≤ 0.05); C, control; G, GEN; D, DEHP; G + D, GEN + DEHP.