Fig. 3

Overexpression of Zc3h8 in COMMA-D cells leads to increased aggressive behavior. a) Mouse mammary COMMA-D cells were stably transfected with empty vector or vector driving Zc3h8 under control of the EF6 promoter and processed for western blot. Images were quantitated using Image J, and results of RT-qPCR are also shown. b) COMMA-D cell lines were grown in 96-well plates and tested for proliferation using luciferase viability assay. ANOVA was used to determine significance p < 0.05. c) Wound healing assay using stably transfected COMMA-D cell lines. ANOVA was used to assess significance at p < 0.01. d) and e) Transwell invasion assay and quantification with COMMA-D stable cell lines. Experiments were performed in triplicate, significance determined to be p < 0.05 by Student’s t-test. For all the above experiments error bars represent standard deviation, and images are representative of typical results (* indicates p < 0.05 and ** indicates p < 0.01). f) BALB/c mice were injected with syngeneic COMMA-D cells that had been transfected with a control vector or one driving expression of ZC3H8 and were monitored for the appearance of tumors for 10 weeks