Figure 3.

CircPRMT5 promoted CRC by reducing the inhibitory effect of miR‐377 by sponge activity. (A). Protein expression of CDK2 and cyclinD1 was determined by Western blotting; (B). mRNA expression of CDK2 and cyclinD1 was determined by RT‐qPCR. Data are mean ± SD. *P < .05; (C). Ago2 RNA immunoprecipitation (RIP) assay for the amount of circPRM5 in CRC cells expressing Flag‐AGO2 or Flag‐GFP. Data are mean ± SD. *P < .05; (D). Luciferase reporter assay for the luciferase activity of LUC‐circPRMT5 or LUC‐circPRMT5‐mutant in HCT‐116 cells cotransfected with miR‐377 mimics. Data are mean ± SD. *P < .05; (E). RNA pull down assay for the amount of circPRMT5 and miR‐377 with circPRMT5 probe in SW620 cells expressing either control or circPRMT5 shRNA or in HCT‐116 cells expressing either with control vector or circPRMT5 overexpression plasmid. Data are mean ± SD. *P < .05; (F). CCK‐8 assay of SW620 cells transfected with the shcircPRMT5 within or without cotransfected with miR‐377 inhibitor. Data are mean ± SD. *P < .05; (G). Colony formation assay of SW620 cells transfected with the shcircPRMT5 within or without cotransfected with miR‐377 inhibitor. Data are mean ± SD. *P < .05, **P < .01; (H). Western blotting analysis of CyclinD1 and CDK2 in SW620 cells transfected with the shcircPRMT5 or HCT‐116 cells expressing with circPRMT5 overexpression plasmid; (I). RT‐qPCR analysis of CyclinD1 and CDK2 in CRC cells transfected with the shcircPRMT5 or with circPRMT5 overexpression plasmid, and within or without cotransfected with miR‐377 mimics or inhibitor. Data are mean ± SD. **P < .01, ***P < .001