Fig 1: Effect of 9cRA on the expression of pro-inflammatory (A-A”) and anti-inflammatory (B-B”) genes and proteins in murine Sertoli cells. Cells were treated with a vehicle (control, C), or 10− 7M 9cRA for 24 h. (A, B) Relative expression of Ifng, Tnfr1, Il1a, Il6, Tgfb1, Il10, Gal1, and Ido mRNAs was determined using quantitative real-time RT–PCR analysis. The histograms are the quantitative representation of data of three independent experiments, each in triplicate. The expression of the individual genes was normalized to the mean expression of the reference genes (Rn18s, B2m, Rpl13a, and Hprt1) as an internal control (relative quantification, RQ). (A’, B’) Western blot detection of TGFβ, IL-10, GAL-1, IDO, IFN-γ, TNFR1, IL-1α, and IL-6 proteins. The histograms are the quantitative representation after densitometry of data (mean ± SD) of three independent experiments, each in triplicate. The relative level of studied protein was normalized to β-actin. The protein levels within the control group were arbitrarily set as 1. Significant differences from control values are denoted as *p < 0.05, **p < 0.01, and ***p < 0.001 (A”,B”) Subcellular localization of TGFβ, IL-10, GAL-1, IDO, IFN-γ, TNFR1, IL-1α, and IL-6 proteins in murine Sertoli cells was visualized by immunofluorescence. Scale bar = 100 μm