Fig. 2

Over-expression of circ-HuR suppresses the growth and aggressiveness of gastric cancer. a MTT colorimetric assay showing the viability of AGS and MKN-45 cells stably transfected with empty vector (circ-Mock), circ-HuR, or lin-HuR. b and c Soft agar (b) and matrigel invasion (c) assays indicating the in vitro growth and invasion of AGS and MKN-45 cells stably transfected with circ-Mock, circ-HuR, or lin-HuR. d Representative (left panel), in vivo growth curve (middle panel), and weight at the end points (right panel) of xenograft tumors formed by subcutaneous injection of AGS cells stably transfected with circ-Mock or circ-HuR into the dorsal flanks of nude mice (n = 5 for each group). e Representative images (left panel) and quantification (right panel) of immunohistochemical staining showing the expression of Ki-67 and CD31 within xenograft tumors formed by hypodermic injection of AGS cells stably transfected with circ-Mock or circ-HuR (n = 5 for each group). Scale bars: 50 μm. f-h Representative images (f), H&E staining (g, arrowheads), and quantification (h, left panel) of lung metastatic colonization and Kaplan-Meier curves (h, right panel) of nude mice treated with tail vein injection of AGS cells stably transfected with mock or circ-HuR (n = 5 for each group). Scale bar: 100 μm. ANOVA and Student’s t-test analyzed the difference in a-e and h. Log-rank test for survival comparison in (h). *P < 0.01 vs. circ-Mock. Data are shown as mean ± SEM (error bars) and representative of three independent experiments in (a-c)