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. 2019 Mar 18;145(4):1068–1082. doi: 10.1002/ijc.32207

Figure 5.

Figure 5

P2X7 activates Wnt/β‐catenin and mTOR/HIF1α/VEGF signaling. (a) mRNA and (d) protein in BzATP‐treated (5, 25 or 125 μM for 24 h) HOS/MNNG cells. (b) mRNA and (e) protein of β‐catenin and TCF1, and (g) immunofluorescent staining for β‐catenin in HOS/MNNG cells treated with BzATP (125 μM) or A740003 (5 μM) for 24 h. (c) mRNA and (f) protein of β‐catenin and TCF1, and (g) immunofluorescent staining for β‐catenin in HOS/MNNG cells transfected with scrambled or shP2X7 lentiviral vectors or treated with shP2X7 and BzATP (125 μM). Expression of total mTOR, phosphorylated (p)‐mTOR (Ser2448) and HIF‐1α in HOS/MNNG cells treated with (i) BzATP (5, 25 or 125 μM) for 24 h, (j) BzATP (125 μM) or A740003 (5 μM) for 24 h, or (k) lentiviral vectors or treated with shP2X7 and BzATP (125 μM). (l) VEGF protein in HOS/MNNG cells treated with BzATP (5, 25 or 125 μM) or A740003 (5 μM) or both for 24 h, or transfected with lentiviral vectors or treated with shP2X7 and BzATP (125 μM). Relative gene or protein expression was assayed by normalizing with Lamin B1 or β‐actin. In Immunofluorescence, secondary antibodies were FITC or Cy3 labeled, and cell nuclei were counterstained with DAPI. VEGF protein was measured using a commercially available human VEGF ELISA kit. Data are means ± SD of 3 independent experiments. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. control. [Color figure can be viewed at wileyonlinelibrary.com]