Suppression of JAM-A expression in H1299 and A549 cells.
(A) Western blot analyses of JAM-A depletion efficiency in cancer cells. Data are shown as representative Western blots (left panel). Densitometry value analysis of 3 independent Western blot experiments, data were normalized against GAPDH, Columns, mean; bars, SD, **p<0.01(right panel). (B) Real-time PCR analyses of JAM-A depletion efficiency in cancer cells, The relative quantity of JAM-A mRNA, normalized to β-actin, were compared to the negative control siRNA-transfected group based on the equation RQ = 2−ΔΔCt. Columns, mean of RQ in a representative experiment; bars, max/min RQ. Experiments were independently repeated 3 times in triplicate with similar results. (C) Flow cytometry assay analyses of JAM-A surface expression depletion efficiency. H1299 and A549 cells were transiently transected with negative control siRNA or JAM-A siRNA; after 48 h, cells were trypsinized and flow cytometry assay analyses were conducted as described in the Methods section. Data are shown as representative histograms (upper panel), mean fluorescence intensity of triplicate values in a representative experiment, Columns, mean; bars, SD. **p<0.01 (lower panel). Experiments were independently repeated 3 times in triplicate with similar results.