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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Elevated Na/H exchanger 1 (SLC9A1) emerges as a marker for tumorigenesis and prognosis in gliomas

Fig. 4

Pharmacological inhibition of NHE1 protein decreases tumor volume, invasion, and angiogenesis. a Experimental protocol. SB28-GFP cells (40,000) or GL26 cells (100,000) were injected in the right striatum of C57BL/6 J mice. Starting 5 days after implantation (d.p.i.), mice received either vehicle (Veh, 1.25% DMSO in PBS, 10 ml/kg/day) or HOE (0.5 mg/kg/day, ip) for 5 consecutive days. Mice were sacrificed at 10 d.p.i. for immunostaining and at 15 d.p.i. for flow cytometry. b Representative immunostaining of fixed brain sections (25 μm) for NHE1 protein expression in SB28-GFP tumor. Data are means ± SEM from four independent experiments. **, p < 0.01. c Shown SB28-GFP tumors of Vehicle group and HOE group at 10 d.p.i., respectively. Arrows, the farthest isolated GFP-positive glioma cells; Dashed line, bulk tumor area border. d HOE treatment decreased SB28-GFP tumor volumes at 10 d.p.i. Data are means ± SD from six independent experiments. **, p < 0.01. e, f Tumor invasion was quantified by measuring the maximal invasion distance, as well as the invasive front as a percentage of total tumor. Data are means ± SEM from six independent experiments. ***, p < 0.005. g Representative confocal images of SB28-GFP tumor sections (25 μm) immunostained for NHE1 and endothelial cell marker CD31. Arrows: NHE1 expression in tumor cells, arrowhead: NHE1 expression in the CD31+ vessels. h Data summary by vessel area/total area. Data are means ± SEM from four independent experiments. *, p < 0.05. i NHE1 expression in vessel area. Data are means ± SEM from four independent experiments. ***, p < 0.001. j Regression correlation analysis of NHE1 expression in vessel area and CD31 expression in tumor. k Representative confocal images of ECM protein laminin staining in SB28-GFP tumor. Arrows: increased accumulation of laminin; Arrowheads: less laminin accumulation. l Summary data of laminin fluorescence intensity. Data are mean ± SEM, from seven independent experiments. *, p < 0.05

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