Silencing SF1 and SREBP1 suppresses DGKθ gene expression. A: H295R wild-type and SF1 knockdown (k.d.) cells were treated with 0.4 mM Bt2cAMP for 48 h and cell lysates were harvested and analyzed by SDS-PAGE (8%), followed by Western blotting for DGKθ, SF1, SREBP1, and GAPDH. B: Real time RT-PCR was used to assess the mRNA expression of DGKθ, SREBP1, and SF1 using total RNA that was isolated from wild-type and SF1 knockdown H295R. Data are graphed as fold change in DGKθ, SREBP1, or SF1 expression mRNA expression normalized to the mRNA expression of β-actin and represent the mean ± SEM of three separate experiments, each performed in triplicate. *Statistically different from untreated control group, P < 0.05. C: Wild-type and SREBP1 knockdown cells were treated with 0.4 mM Bt2cAMP for 48 h and cell lysates were harvested and analyzed by SDS-PAGE and Western blotting for DGKθ, SF1, SREBP1, and GAPDH. D: RNA isolated from wild-type and SREBP1 knockdown H295R cells was subjected to qRT-CPR analysis. Data are graphed as fold change in DGKθ, SREBP1, or SF1 expression mRNA expression normalized to the mRNA expression of β-actin and represent the mean ± SEM of three separate experiments, each performed in triplicate. *Statistically different from untreated control group, P < 0.05.