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

Figure 3 Selection (A, B) and inducibility (C, D) of stably-transfected Sβtet29Cap clones. The stable Sβtet29 clone was transfected with the pTRE2hygCapG full size vector and clones selected with hygromycin B (200 μg/mL). A: Individual clones (n = 49) were induced with 500 ng/mL doxycycline for 24 h and the protein lysate subjected to Western blotting. The CapG level was calculated for each individual clone in the non-induced and the induced state. The fold increase (normalised to actin) in CapG for each of the 49 clones is shown in the bar chart; B: A representative Western blotting for CapG is shown for pTRE2hygCapG clones 30 to 35. The indicates the basal CapG expression, + doxycycline (dox) induced; C: The inducibility of CapG protein expression was investigated by treating the Sβtet29Cap35 cells with 0, 1, 5, 10, 50, 100, 500, 1000 ng/mL dox for 24 h. The resulting Western blotting for CapG and GAPDH as a loading control is shown; D: The Western blotting represents the CapG protein level of Sβtet29Cap35 cells, induced with 500 ng/mL doxycycline for 0, 12, 24, 36 and 48 h. GAPDH was used as a loading control.