Additional file 1: of Regulation of the oncogenic phenotype by the nuclear body protein ZC3H8

Figure S1. Reduced expression of Zc3h8 in a second cell line and using a second targeting shRNA leads to reduced invasive behavior. A) Western blot demonstrating reduced levels of ZC3H8 protein in cV1A 03–31 cells transfected with a vector driving expression of an shRNA targeting site A. B) cV1A 03–31 cells transfected with a vector driving expression of siRNA targeting Zc3h8 at site A close a wound more slowly than cells transfected with a negative control vector in vitro. ANOVA revealed a significant difference of p < 0.05. C) A second tumor cell line, cV1A 01–51, was transfected with a vector targeting Zc3h8 at site C or negative control. Cells with reduced Zc3h8 closed a wound more slowly than negative control cells. ANOVA was used to assess significance at 12 h of p < 0.001. D) cV1A 01–51 cells with targeted Zc3h8 also formed smaller growths in syngeneic BALB/c mice than negative control cells. Student’s t-test was used to determine p < 0.001. E) cV1A 01–51 cells with reduced Zc3h8 did not form colonies in soft agar, while those transfected with a negative control formed large growths after two weeks. F) Tumor cell lines were surveyed by RT-qPCR for relative expression of Zc3h8 and Gata-3 compared to levels in virgin mammary gland, shown in a scatter plot. G) Overexpression of Zc3h8 in COMMA-D cells did not lead to a decrease in Gata-3 levels as determined by RT-qPCR. Error bars represent standard deviation. (TIF 22335 kb