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. 2016 Jan 19;17(1):117. doi: 10.3390/ijms17010117

Figure 6.

Figure 6

Figure 6

CHR20/CHR21 increase phosphorylation levels in PI3K/Akt and MEK/ERK1/2. RGC-5 cells were treated with 10 μM CHR20 and CHR21, repectively for 5–80 min or 0.3–30 μM CHR20/CHR21 for 40 min. By applying Western blotting with antibodies including anti-phospho-Akt (Ser473), anti-phospho-ERK1/2, anti-phospho-and eNOS (Ser1177), respectively, the phosphorylation of the relevant proteins were determined. (A) CHR20 and CHR21 time-dependently increased the phosphorylation levels of Akt and ERK1/2 in RGC-5 cells; (B) CHR20 and CHR21 dose-dependently induced the phosphorylation of Akt and ERK1/2 in RGC-5 cells; (C) Effects of pathway inhibitors on the phosphorylation of Akt, eNOS and ERK1/2 affected by CHR21. The density of the blot in each lane was presented as mean ± standard deviation. Each data was calculated based on at least three individual experiments. * p < 0.05, ** p < 0.01 vs. control group. # p < 0.05, ## p < 0.01 vs. CHR21 pre-treated group. Blots were quantified using ImageJ software. pd: PD98059; ly: LY294002; p-eNOS: phosphorylated protein level of eNOS; pAkt: phosphorylated protein level of Akt; pErk: phosphorylated protein level of ERK; CHR: abbreviation of Chen HeRu; T-Erk: total protein level of ERK; p-NOS: phosphorylated protein level of NOS; T-NOS: total protein level of NOS.