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Fig. 5 | Molecular Cancer

Fig. 5

From: Long noncoding RNA GAS5 inhibits progression of colorectal cancer by interacting with and triggering YAP phosphorylation and degradation and is negatively regulated by the m6A reader YTHDF3

Fig. 5

LncRNA GAS5 inhibits colorectal cancer progression via attenuation of YAP-mediated expression of YTHDF3 in vitro and in vivo. a-b CCK8 proliferation assays were performed to determine cell proliferation of CRC cells after co-transfection of GAS5 and YTHDF3 plasmid (a) or GAS5 and YTHDF3 specific shRNAs (b). The mean ± SD is shown for five independent experiments. ***P < 0.001. c-d The scratch wound-healing assay were performed to investigate the changes in migratory abilities of HCT116 and LOVO cells after indicated transfection, respectively. The wound-healing percentage was analyzed with the ImageJ software (right). All experiments were performed in triplicate, and results are presented as mean ± SD. ***P < 0.001. e Transwell analysis assays were performed in the indicated CRC cells after transfection, respectively. Transwell analysis assays were quantified using the ImageJ software (right). All experiments were performed in triplicate, and results are presented as mean ± SD. ***P < 0.001 (f) Representative images of tumors growth in xenografted BALB/c nude mice. Each group of mice were ectopically implanted with 2 × 106 indicated cells into the flanks of mice (n = 6). Here, cells were transfected with indicated lentiviral vector. Mice were sacrificed 25 days after implantation and tumor volume was measured (error bars represent mean ± SD). Scale bar, 1 cm. ***P < 0.001. g Representative images of ISH and IHC staining on paraffin-embedded samples of xenograft tumors growth in BALB/c nude mice. h Representative lung tissues images of lung metastasis model generated by injecting tumor cells into the tail veins of mice. HE staining showing the number and foci of lung metastases in each group (n = 6); **P < 0.01. And the area of metastases nodules in individual mice was calculated using Dmetrix software (bottom panel)

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