Figure 2. The effect of NMB on p65, c-Jun, COX-2 and IL-6 expression in cultured primary myometrial cells.

Cells were treated with 1 μM NMB for 0, 0.5, 1, 2, 4, and 6 h, and the protein levels were analyzed using Western blotting or ELISA, and the mRNA levels were analyzed by qRT-PCR. (A) Expression of total p65, p-p65 (Ser⁵³⁶), total c-Jun, p-c-Jun (Ser⁶³, Ser⁷³) and COX-2 was analyzed using Western blotting with GAPDH as a control standard. (B) IL-6 in the supernatants was quantified by ELISA. (C) The relative level of the COX-2 protein was normalized to that of the internal control (GAPDH). The relative levels of phospho-p65, and phospho-c-Jun protein were normalized to total p65 or c-Jun, respectively. (D) IL6 and PTGS2 mRNA levels were analyzed by qRT-PCR. Blots are representative. Data are expressed as mean ± SD of three independent experiments (n=3). *P<0.05; **P<0.01, against the control group.