Fig. 3

IL30-dependent regulation of IGF1 production in human PC cells and IGF1-mediated autocrine growth loop. A, B Elisa assay of IGF1 release by wild-type, EV and IL30 gene-transfected DU145 (A) and PC3 (B) cells. ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with WT and EV-transfected cells. Results are expressed as mean ± SD. C Cytofluorimetric analyses of IGF1R expression in PC3 and DU145 cells. Red lines: isotype control. Experiments were performed in triplicate. D, E Elisa assay of IGF1 release by wild-type DU145 (D) and PC3 (E) cells, after the treatment with anti-IL30 Abs. (D) ANOVA: p < 0.05. *p < 0.05, Tukey HSD test compared with DU145 cells untreated or treated with 5 μg/mL. E ANOVA: p < 0.001. *p < 0.01, Tukey HSD test compared with untreated PC3 cells. Results are expressed as mean ± SD. F, G MTT assay of DU145 (F) and PC3 (G) cells, untreated (0.0 ng/mL) or treated with rhIGF1 (5.0, 10, 30, 50 ng/mL). ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with 0 ng/mL. **p < 0.01, Tukey HSD test compared with 0 and 5 ng/mL. ***p < 0.01, Tukey HSD test compared with 0, 5 and 10 ng/mL. Results are expressed as mean ± SD. H, I MTT assay of DU145 (H) and PC3 (I) cells, untreated (0.0 μg/mL) or treated with anti-IGF1 Abs (0.1, 0.4, 0.8 μg/mL in DU145; 0.25, 0.50, 0.70 μg/mL in PC3). (H) ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with 0.0 μg/mL. **p < 0.01, Tukey HSD test compared with 0.0 and 0.1 μg/mL. ***p < 0.05, Tukey HSD test compared with 0.0, 0.1 and 0.4 μg/mL. (I) ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with 0.00 µg/mL. **p < 0.01, Tukey HSD test compared with 0.00 and 0.25 μg/mL. Results are expressed as mean ± SD. J, K MTT assay of wild-type and IL30 gene-transfected DU145 (J) and PC3 (K) cells, untreated (0.0 μg/mL), or treated with anti-IGF1 Abs (30 μg/mL). ANOVA: p < 0.0001. *p < 0.01, Tukey HSD test compared with wild-type cells. **p < 0.01, Tukey HSD test compared with wild-type and IL30-transfected cells. Results are expressed as mean ± SD