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. 2021 Feb 8;6(3):e142299. doi: 10.1172/jci.insight.142299

Figure 6. NGBR knockdown attenuates growth factor–induced phosphorylation of AKT and ERK1/2 in HemSCs.

Figure 6

Protein levels were determined by Western blot. NGBR knockdown reduced the FGF2-induced (A), VEGF-induced (B), and EGF-induced (C) phosphorylation of AKT and ERK1/2 in HemSCs. Twenty-four hours after siControl or siNGBR transfection, cells were arrested overnight in serum-free medium and then stimulated with FGF2 (100 ng/mL), VEGF (100 ng/mL), and EGF (100 ng/mL) in serum-free medium at indicated times (5, 10, 20, and 30 minutes). The growth factor–induced phosphorylation AKT and ERK1/2 were determined by Western blot. Total AKT, total ERK1/2, and actin protein levels were used as respective loading controls. Quantitative analysis of phosphorylated proteins was carried out using ImageJ software, and proteins were normalized to total proteins correspondingly. *P < 0.05 vs. control (siControl) cells. #P < 0.05 vs. control (siControl) cells treated by FGF2, VEGF, and EGF (n = 3), 3 repeats. Statistical analyses: 1-way ANOVA with Bonferroni’s post hoc test; data are expressed as mean ± SEM. NGBR, NOGOB receptor; HemSCs, hemangioma stem cells; siControl, control siRNA; siNGBR, NGBR siRNA.