Figure 5
Confirmation of materials and adequate time to influence RIPK1 and RIPK3 mRNA and expression effects of TNF and IFNG on the expression of RIPK1 and RIPK3 protein in cultured luteal steroidogenic cells (LSCs).
LSCs were treated with TNF (2.3 nM) and/or IFNG (2.5 nM), PGF (1.0 μM) or NONOate (100 μM) for 12 and 24 h. (A) and (C) show the expression of RIPK1 mRNA, and (B) and (D) show the expression of RIPK3 in the LSCs after culture for 12 and 24 h, respectively. Data are expressed as the relative ratio of RIPK1 or RIPK3 mRNA to GAPDH mRNA levels. All values are the means ± SEM. All of the experiments were repeated more than three times. The data were statistically analyzed by ANOVA followed by Dunnett’s multiple comparison test. Asterisks indicate significant differences compared with control (*P < 0.05, ***P < 0.01, respectively). (E) and (F) show representative western blot bands for RIPK1 or RIPK3 and ACTB in cells treated with TNF (2.3 nM) and IFNG (2.5 nM) for 24 h. The resulting signal was visualized using the alkaline phosphatase visualization procedure and quantitated by computer-assisted densitometry. Data are expressed as the relative ratio of RIPK1 or RIPK3 protein to ACTB protein. All values are the means ± SEM. All of the experiments were repeated more than three times. The data were statistically analyzed by Student’s t-test. Asterisks indicate significant differences compared with control for 24 h (P < 0.05).
