Figure 3.
Prep1 overexpression increases Bcl-XL expression (A; left) Total RNA was purified from untreated Prep1-overexpressing or control F9 cells, retrotranscribed using polyA+ primers and semiquantitative PCR analysis performed with specific primers for murine Bcl-XL and β-actin. The results of the densitometric analysis is shown under each lane. (Right) Crude extracts from the above cells were resolved by 12% SDS-PAGE and transferred to PVDF membrane. Endogenous Bcl-XL protein levels were analyzed by immunoblotting with specific monoclonal antibody and β-actin was used for normalization. Results of the densitometric analysis are shown under each lane. (B) Total mRNA and crude extracts were prepared and processed as above for qPCR (experiment performed in triplicate; left) or immunoblotting (right) from cell that were treated (or not) with the etoposide for 12 h. A value of 1 and of 100 was arbitrarily given to Bcl-XL mRNA (left) and protein (right) amount, respectively, in untreated cells infected with the control vector.