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. 2018 Jan 5;32(3):1170–1183. doi: 10.1096/fj.201700834R

Figure 5.

Figure 5.

Effect of growth at low pH and of GPR68 on gene expression and IL-6 expression by CAFs and PDAC cell proliferation. A) Real-time qPCR analysis of expression of fibrosis-related genes in CAFs cultured in pH 7.4 or 6.8 medium for 6, 12, and 24 h. Expression was normalized to that at 0 h. Data are means ± sem, n = 3, unpaired Student’s t test. **P < 0.01, ***P < 0.001, ****P < 0.0001. B) CAFs transfected with control or GPR68 siRNA were cultured in pH 7.4 or 6.8 medium for 6 h. IL-6 expression was measured by real-time qPCR and normalized to expression with control siRNA at pH 7.4. Data are means ± sem, n = 3; 2-way ANOVA, followed by Tukey post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001. C) CAFs transfected with control or GPR68 siRNA were cultured in pH 7.4 or 6.8 medium for 24 and 48 h. IL-6 in CM was assessed by ELISA. Data are means ± sem, n = 3; 2-way ANOVA, followed by Tukey post hoc test. ****P < 0.0001. D) CEllTiter-Glo luminescent (proliferation) assay of BxPC cells cultured in CM from CAFs transfected with control or GPR68 siRNA for 72 h with or without IL-6 neutralizing antibody (ab). Data are means ± sem, n = 4; 2-way ANOVA, followed by Tukey post hoc test. *P < 0.05, ****P < 0.0001.