Overexpression of AKR1C isoforms 2 and 3 promotes preneoplastic transformation. (A) V5-tagged AKR1C isoforms 2 and 3 were overexpressed in senescent NHEKs from adenoviral vectors. Senescent NHEKs were infected with Ad-GFP, Ad-AKR1C2-V5 (Ad-C2-V5) or Ad-AKR1C3-V5 (Ad-C3-V5) particles, and analyzed 48 h later by western blot using an-anti V5 antibody and an anti-GAPDH used as loading control. (B) Once infected as in (A), the emergence frequency was determined as in Figure 5D. Countings were performed in triplicate (Student t-test; *p < 0.05, of the comparison of the control-GFP condition with Ad-C2-V5/Ad-C3-V5 conditions).