Tetracycline-inducible protein expression in pancreatic cancer cells: Effects of CapG overexpression

Figure 1 Identification of clones transfected with CMV-rtTA2S-M2 or pN1βactin-rtTA2S-M2-IRES-EGFP-SUIT-2 cell clones. Suit-2 cells were transfected with CMV-rtTA2S-M2 (A) or pN1βactin-rtTA2S-M2-IRES-EGFP (B) and stable clones selected using 300 μg/mL G418. Clones were isolated and transfected with pTRE2hygluc, used as an indirect measure of rtTA activity. Cells were treated for 24 h with 500 ng/mL doxycycline (black bars) or an equivalent volume of PBS (white bars), and luciferase activity measured in 50 μg protein (1 mg/mL protein; 50 μL was assayed per sample). Error bars represent standard errors for three experiments. C: Cells were transiently transfected with pTRE2hygGST-P, pTRE2hygCYP2E1, and pTRE2hygS100A6 and treated with 500 ng/mL doxycycline or PBS as control for 24 h. Protein lysates were prepared and subjected to Western Blotting using anti-GST-P, CYP2E1 and S100A6 antibodies. α-tubulin was used as a loading control.