Figure 2

Validation of gene silencing by CHK1 siRNA. MIA PaCa-2 cells were transfected with either CHK1 or control siRNA and allowed to grow for 48–72 hrs. (A) Total RNA from the siRNA treated MIA PaCa-2 cells was isolated at 48 hrs and analyzed by qRT-PCR for CHK1 expression. CHK1 expression for each siRNA treatment was compared to untreated cells. GAPDH was used as an internal control for all the samples and fold change was calculated by normalizing all the data to GAPDH expression. (B) Lysates from CHK1 siRNA treated MIA PaCa-2 cells were prepared at 72 hrs post transfection and analyzed by western blot for expression of CHK1 protein using an anti-CHK1 antibody. (C) CHK1 siRNA treated cells showed decreased growth of MIA PaCa-2 cells at 72 hours after siRNA transfection when compared to no siRNA treatment or non-silencing siRNA treatment. Cell images were taken at 20× magnification.