Fig. 4

ZNF561-AS1 silencing suppresses CRC progression in vitro and in vivo. a ZNF561-AS1 silencing inhibited HCT-116 growth. n = 3, *p < 0.01. b Colony formation assay. ZNF561-AS1- or mock-silenced HCT-116 cells were planted in 6 cm dishes and colonies counted after 12 days. Formed colonies were stained with crystal violet and counted. n = 3, *p < 0.05. c Ki-67 level was significantly reduced in ZNF561-AS1 depleted cells compared to controls assessed by immunofluorescence staining. n = 3, *p < 0.01. d PCNA, Cyclin D1, CDK4, p21, and cleaved caspase-3 levels in ZNF561-AS1- and mock-silenced HCT-116 cells. Actin served as loading control. Representative blots are shown (left panel). Ratios of PCNA, Cyclin D1, CDK4, p21, and cleaved caspase-3 vs Actin levels were computed after densitometric analysis of blot images on Image J (right panel). Data are 3 independent experiments. *p < 0.05. e Knockdown of ZNF561-AS1 induced cell apoptosis assessed by AO/EB staining. n = 3, *p < 0.01. f Knockdown of ZNF561-AS1 induced cell apoptosis assessed by TUNLE staining. n = 3. g The expression of ZNF561-AS1, miR-26a-5p and miR-128-3p in xenograft tumors. n = 3, *p < 0.05. h Levels of SRSF6 in xenograft tumors. Actin was served as loading control. Representative blots were shown (left penal). Ratios of level of SRSF6 vs Actin were calculated after densitometric analysis of blot images using NIH Image J 1.61 (right panel). Data were from three individual experiments. *p < 0.01. i Knockdown of ZNF561-AS1 reduced xenograft tumors weight. n = 3, *p < 0.05. j Knockdown of ZNF561-AS1 reduced xenograft tumors volume. n = 3, *p < 0.05. k Ki-67 immunohistochemical staining in xenograft tumor specimens. n = 3, *p < 0.05