Figure 1

Time-course reduction in PKCε mRNA expression after transfection of luteal steroidogenic cells with PKCε specific siRNA. (A) Representative RT-PCR products obtained from total RNA using the PKCε and GAPDH primers. The amount of total RNA was adjusted to 200 ng per reaction and 40 cycles were used for PKCε ; while 28 cycles were used for GAPDH. The size of the amplified products for the GAPDH and PKCε were 900 and 500 bp, respectively. PKCε and GAPDH mRNA expression after 48, 72, and 96 h of transfection with PKCε specific siRNA are shown. Lanes labeled Media, non-specific (Non-sp) siRNA, and Transfection reagent represent respective treatments without PKCε specific siRNA treatment. GAPDH was used as the control gene to normalize the PKCε mRNA expression. (B) Quantitative analysis of the RT-PCR products obtained in four (n = 4) replicates similar to those shown in panel A. Data are the mean mean ± SEM of the densitometry measurements for PKCε relative to GAPDH mRNA. Statistical comparisons were made between different treatments. Different letters above each SEM represent different values (P < 0.05).