Figure 4.

BJ induced apoptosis and reduced EGFR and pEGFR^Y1068 levels in H1975 cells. (A) Western blot analysis. The protein lysates from H1975 cells as treated with 1, 2, and 5 mg/mL of BJ extract for 12 hours were collected and used for Western blot analysis. The blots were incubated with various primary antibodies, including EGFR, phosphorylated EGFR^Y1068, Akt, phosphorylated Akt^S473, caspase-3, and PARP as specified, which were followed by HRP-conjugated secondary antibodies. GAPDH was used as loading control. The blots were visualized by ECL detection system. (B) Densitometric determination of EGFR and phosphorylated EGFR^Y1068 amelioration. The densitometric ratios of EGFR and phosphorylated EGFR^Y1068 in H1975 cells from Western blot analysis were obtained by first normalizing individual band intensity at each concentration to that of the loading control and compared with those of water treatment. The results were expressed as mean values of three independent experiments (*P<0.05, **P<0.01, unpaired Student’s t-test as compared with control water).

Abbreviations: BJ, Brucea javanica; EGFR, epidermal growth factor receptor; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; PARP, poly(adenosine diphosphate ribose) polymerase; pEGFR, phosphorylated EGFR.