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. 2017 Mar 6;8(20):32655–32667. doi: 10.18632/oncotarget.15948

Figure 5. AG1478 and NAC attenuate HG-induced EGFR signaling activation, ROS generation, endoplasmic reticulum stress, cell firosis and apoptosis in SV40 cells.

Figure 5

SV40 cells pretreated with AG1478 (10 μM) or NAC (10 mM) for 1 h were incubated with HG (33 mM) for 1 h. Then cells were lysed and the extracted total proteins were processed for the detection of p-EGFR and p-AKT using Western blot; and statistic figure was shown, data were presented as mean ± SDs. (Six mice in each group were used for above analysis (A). (B) AG1478 and NAC inhibit high glucose-induced ROS generation. SV40 cells pretreated with AG1478 (10 μM) or NAC (10mM) for 1h were incubated with HG (33 mM) for 2h. DCFH-DA probes were loaded and the ROS positive cells were detected using the fluorescence microscope. Also, after loading with the probes, cells were processed to flow cytometry analysis for O2 level, and mean fluorescence intensity (MFI) value was determined. Data were presented as mean ± SDs. (n = 3 for each experiment. **P < 0.01, ***P < 0.001 versus HG; ##P < 0.01, ###P < 0.001 versus DMSO). (C) Western blot analysis for the protein expression of ERS-related proteins ATF4 and CHOP in SV40 cells. (D) Western blot analysis for the fibrosis and apoptosis protein expression of TGF-β1 and Cleaved-Caspase 3 in SV40 cells.