Figure 4

Verification of pBLIC-Bax, pBLIC-catalase or pBLIC-p53 transduction and protein overexpression. Either LNCaP or H358 cells were transduced with the indicated overexpression constructs and selected in G418-containing media as described in the Materials and Methods section, and harvested within 7 days of selection. RT-PCR and Western blot analysis was used to confirm overexpression of mRNA and protein from the constructs. RT-PCR and Western blotting was carried out on RNA and protein samples from cells transduced with the indicated constructs. Cells transduced with the empty pBLIC-Neo vector were used to determine baseline levels of the two proteins. Single bands were seen corresponding to the full-length mRNA (top panel, RT-PRCR) or protein (bottom panel, Western blotting) in all cases. GAPDH is shown as a loading control for both RT-PCR and Western blotting. (A) Overexpression of Bax mRNA and protein from pBLIC-Bax in LNCaP cells. (B) Overexpression of catalase mRNA and protein from pBLIC-catalase in LNCaP cells. (C) Overexpression of p53 mRNA in LNCaP cells and p53 protein in H358 cells from the pBLIC-p53 construct.