Sc 166821

All cell culture materials were obtained from Gibco Inc. Recombinant Human Activin-A (GFH6) was purchased from Cell Guidance Systems. RepSox (#72392), a cell permeable, selective inhibitor of the TGF-β type 1 receptor (TGFβRI) ALK5 was obtained from Stemcell Technologies. Recombinant forms of FSH (Gonal-F) and hCG (Ovitrelle) was purchased from Merck Global (Darmstadt, Germany). SAPK/JNK Kinase Assay Kit (#8794, nonradioactive), Hoechst 33342 (#4082), anti c-Jun (#9165), anti-phospho-c-Jun^Ser73 (#3270S), Anti-Phospho-SAPK/JNK^{Thr183/Tyr185} (#9251), Smad2 (#3122) and Phospho-Smad2^{Ser465/Ser467} (#18338) antibodies were obtained from Cell Signaling. Oil Red O was purchased from Sigma Inc. (USA). All western blotting buffers and reagents were purchased from Bio-Rad. Anti-Vinculin Antibody (V9264) was purchased from Sigma-Aldrich. Mouse antihuman monoclonal antibodies were purchased from Santa Cruz Biotechnology for the detection of human 3β-HSD Type II (sc-100466), 17β-HSD type-I (sc-376719), StAR (sc-166821), and P450 SCC (CYP11A1, sc-292456). Aromatase (CYP19A1, ab34193) monoclonal mouse antibody was from Abcam Inc. YO-PRO®-1 Iodide (491/509) was obtained from Life Technologies. Texas Red™-X Phalloidin was obtained from Thermo Fisher Scientific.

Four-micrometre tissue sections from formalin-fixed, paraffin-wax-embedded human endometriosis tissues and endometrial tissues were dewaxed, rehydrated, and subjected to high-temperature antigen retrieval. The tissues were incubated with blocking antibody diluent at room temperature for 2 h, and then incubated overnight at 4 °C with the following primary antibodies: anti-vimentin (mouse, ab8978, dilution 1:1000; Abcam), anti-CXCL8 (rabbit, sc-8427, dilution 1:150; Santa Cruz), anti-C7 (rabbit, ab126786, dilution 1:150; Abcam), anti-S100A10 (rabbit, sc-81153, dilution 1:150; Santa Cruz), anti-C3 (rabbit, ab200999, dilution 1:150; Abcam), and anti-StAR (rabbit, sc-166821, dilution 1:150; Santa Cruz). The slides were then incubated with secondary antibody (HRP polymer, anti-mouse/rabbit IgG, Abcam) at room temperature for 1 h. Nuclei were stained with 4-6-diamidino-2-phenylindole (DAPI, ab104139, Abcam) after all antigens had been labelled and then imaged.