Fig. 2
Functional characterization of protein kinase R (PKR) knockdown in pancreatic cancer cell lines PANC-1 and MIA PaCa-2. (a) Bar graphs representing relative PKR mRNA expression levels in PANC-1 and MIA PaCa-2 cells post-treatment with control siRNA (Control si) and two different PKR siRNAs (PKR si 1 and PKR si 2). The y-axis displays normalized mRNA expression. Data represent the mean ± standard error of the mean of four replicates. *P < 0.05, vs. control, Student’s t-test. (b) Western blot analysis showing the protein levels of PKR in PANC-1 and MIA PaCa-2 cells after siRNA treatment. Samples treated with Control si, PKR si 1, and PKR si 2 are depicted. (c) Representative images of PANC-1 and MIA PaCa-2 cells on day 3 after control or PKR siRNA transfection. (d) Line graphs depicting the proliferation of PANC-1 and MIA PaCa-2 cells, as measured by absorbance at 490 nm, post-siRNA treatment. Data represent the mean ± standard error of the mean of five replicates. *P < 0.05, vs. control, Student’s t-test. (e) Apoptosis assay of PANC-1 cells treated with PKR siRNA, using Annexin V and 7-AAD staining and analyzed using flow cytometry. Dot plots show the proportions of live (Annexin V-negative, 7-AAD-negative), early apoptotic (Annexin V-positive, 7-AAD-negative), and late apoptotic (Annexin V-positive, 7-AAD-positive) cells. (f) Quantitative analysis of apoptotic cell distribution from the data in (e). The bar graph shows the percentages of live along with early and late apoptotic cells. Data represent the mean ± standard error of the mean of three replicates. *P < 0.05, vs. control treatment, Student’s t-test.
