Figure 3.

RNAi reduces kindlin-2 levels. A. Quantitative PCR analysis of C2C12 cells treated with kindlin-2 or scrambled siRNA. Samples for analysis were collected 1 day (GM), 3 days (DM day 2) and 5 days (DM day 4) after transfection. Relative kindlin-2 levels were (si scrambled vs. si mig2): 33% vs. 8% (GM), 100% vs. 28% (DM day 2), and 83% vs. 36% (DM day 4). Numbers reflect percent level as compared to scrambled RNAi at DM day 2. Values were averaged from 4 independent transfections, each analyzed in triplicate. B. Immunoblot of kindlin-2 on protein extracts obtained after 2 days in DM (72 hrs after transfection). A band corresponding to kindlin-2 (arrow) is observed in the scrambled RNAi lane only. Equal loading was confirmed by reprobing the blot with S6 kinase (S6K) and GAPDH (data not shown).