Fig. 3

LncRNA GAS5 inhibits colorectal cancer progression via suppression of YAP in vitro and in vivo. a Co-transfected YAP with GAS5 reversed the GAS5-mediated decrease in CTGF expression in HCT116 and DLD1 cells, examined by western blot. b Knockdown of YAP inhibited the sh-GAS5-mediated up-regulation of CTGF in LOVO and RKO cells, examined by western blot. c-d CCK8 proliferation assays were performed to determine cell proliferation of CRC cells after co-transfection of GAS5 and YAP plasmid (c) or GAS5 and YAP specific shRNAs (d). The mean ± SD is shown for five independent experiments. ***P <0.001. e-f Transwell assays were performed to investigate the changes in invasion abilities of CRC cells transfection, respectively. Transwell assays were quantified using the ImageJ software (right). All experiments were performed in triplicate, and results are presented as mean ± SD. ***P <0.001. g-h Representative images of tumors growth in xenografted BALB/c nude mice. Each group of mice were ectopically implanted with 2 X 10⁶ indicated cells into the flanks of mice (n = 6). Here, cells were transfected with indicated lentiviral vector or inducible shRNA. And the volume of tumors in individual mice was calculated using ImageJ software (right panel). Results are presented as mean ± SD. ***P <0.001. i-j Representative images of ISH and IHC staining on paraffin-embedded samples of xenograft tumors growth in BALB/c nude mice. k-l Representative lung tissues images of mice lung metastasis number and foci are shown by HE staining. And the area of metastases nodules in individual mice was calculated using Dmetrix software (bottom panel). (n = 6); **P <0.01