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. 2015 Oct 15;5(11):3339–3349.

Figure 4.

Figure 4

Phenformin inhibits tumor angiogenesis dependent on KRASG12C. A. Western blot analysis to assay the KRASG12C in empty vector transfected cells and KRASG12C transfected cells. GAPDH was used as a loading control. B. Representative images of hTERT-HME1 cells plated on the CAM. Scale bar: 0.5 cm. Qualitative assessment of angiogenesis in the CAM assay. Data are from three independent experiments and are mean ± SD. N = 6, **P < 0.01 compared with wild type cells, ##P < 0.01 compared with KRASG12C knock-in cells. C. KRASG12C up-regulates the expression of pro-angiogenic factors in KRASG12C knock-in hTERT-HME1 cells. Gene expression analysis was performed by real-time PCR comparing parental hTERT-HME1 with KRASG12C clone. Data are from three independent experiments and are mean ± SD. N = 6, *P < 0.05, **P < 0.01 compared with wild type cells and #P < 0.05, ##P < 0.01 compared with KRASG12C knock-in cells. D. Quantification of secreted VEGFA in hTERT-HME1 cells by ELISA. Data are from three independent experiments and are mean ± SD. N = 3, **P < 0.01 compared with wild type cells, ##P < 0.01 compared with KRASG12C knock-in cells. E. Invasion of HUVECs was enhanced by the supernatant of hTERT-HME1 KRASG12C knock-in cells compared with the wild type counterpart. Scale bar represents 50 μm. Data are from three independent experiments and are mean ± SD. N = 3, **P < 0.01 compared with supernatant from wild type cells, ##P < 0.01 compared with supernatant from KRASG12C knock-in cells.