Figure 4.

ROS stimulated MAPK signaling pathway activation in clear cell renal cell carcinoma cells. (A) DHE-ROS analysis by flow cytometry and (B) quantitative analysis of ROS level in 786-O cells following treatment with the ROS scavenger NAC (20 mM, 24 h) or ROS generator H₂O₂ (1 mM, 2 h). (C-E) Western blotting of (C) p-ERK, ERK and p-ERK/ERK, (D) p-p38, p38 and p-p38/p38 and (E) p-JNK, JNK and p-JNK/JNK in 786-O cells following treatment with NAC or H₂O₂. (F) mRNA expression and (G) protein expression of MMP2 in 786-O cells following treatment with NAC or H₂O₂. U6 or GAPDH was used as an internal control. Each analysis was performed at least three times and each sample assessed in triplicate. Data were expressed as the means ± standard deviation. ns, Non-significance; ^*P<0.05, ^**P<0.01 and ^***P<0.001 vs. control. ns, not significant; DHE, dihydroethidium; MMP2, matrix metalloproteinase 2; NAC, N-acetylcysteine; ns, not significant; p, phosphorylated; ROS, reactive oxygen species.