Involvement of the ER subtype on the E2-mediated inhibition of the FC-induced NOS2 up-regulation.

<p>(a) Pre-transfection of CEC with ERα siRNA did not significantly affect the E2-mediated inhibition on the FC-induced increases of the levels of NOS2 protein. (b) Pre-transfection of CEC with ERβ siRNA abolished the E2-mediated inhibition on the FC-induced increases of the levels of NOS2 protein. Top panel: representative results of NOS2 and G3PDH protein levels determined by Western blot analysis. Membrane was probed with anti-G3PDH antibody to verify equivalent loading. Bottom panel: quantitative results of NOS2 protein levels, which were adjusted with G3PDH protein level and expressed as fold-induction of its own control. Values represent the means±s.e.mean. (n = 3). ^*<i>P</i><0.05, different from cells treated with FC. (c) Pre-transfection of CEC with ERα siRNA did not significantly affect the E2-mediated inhibition on the FC-induced increases of the levels of NOS2 mRNA. (d) Pre-transfection of CEC with ERβ siRNA abolished the E2-mediated inhibition on the FC-induced increases of the levels of NOS2 mRNA. The levels of mRNA were detected by RT-PCR (top panel) or real-time quantitative PCR (bottom panel). ER, estrogen receptor; N-T Si, non-target siRNA; Si, small interfering RNA.