Research Paper Volume 13, Issue 22 pp 24605—24620

Figure 6. The protective effect of TRPM2 knockdown in DN by blocking TGF-β1-activated JNK1 activation. (A) Protein assay of TRPM2 and TGF-β1 expression in HK-2 cells stimulated with HG for 24 h. (B) Protein analysis was utilized to confirm the efficacy of TRPM2 silencing in HK-2 cells transfected with si-TRPM2. (C) HK-2 cells were instantly transfected with si-TRPM2 or Si-NC for 48 h, followed by NG or HG treatment for another 24 h. The protein levels of TGF-β1 and p-JNK1 were detected by protein assay. (D) HK-2 cells were instantly transfected with si-TRPM2 or Si-NC for 48 h, followed by NG or HG treatment for another 24 h. In another experimental group, HK-2 cells were incubated with 10 μM SP600125 (a JNK inhibitor) for 1 h, followed by HG treatment for 24 h. Protein assay of CTGF, α-SMA, FN, Collagen I and Collagen III expression in HK-2 cells. (E) qRT-PCR analysis of IL-1β, IL-6, IFN-α, TNF-α and MCP-1 expression in HK-2 cells. (F) Protein assay of TGF-β1, p-IKKα and p-NF-κB expression in HK2 cells. (G) Protein assay of the NF-κB levels in the nuclei of HK2 cells. (H) The protective effect of TRPM2 knockdown in DN by blocking TGF-β1-activated JNK1 activation. The data are shown as the mean ± SD of three independent experiments; *p < 0.05, **p < 0.01 and ***p < 0.001.