Figure 4. SOCS2-AS1 promoted SOCS2 expression through sponging miR-1264. (A) qRT-PCR analysis for indicated genes. (B) Expression correlation between SOCS2-AS1 and SOCS2 in CRC tissues according to TCGA data. GEPIA tool (http://gepia.cancer-pku.cn/detail.php) was used to analyze TCGA data. (C) qRT-PCR was used to expression correlation between SOCS2-AS1 and SOCS2 in CRC tissues. (D) Subcellular location of SOCS2-AS1 was determined by qRT-PCR. (E) Predicted binding sites among SOCS2-AS1, miR-1264 and SOCS2 through miRDB tool (http://mirdb.org/). (F, G) Luciferase reporter assays were carried out to validate the interactions among SOCS2-AS1, miR-1264 and SOCS2. (H) RNA pulldown assay was conducted to confirm the interaction between SOCS2-AS1 and miR-1264. (I) RIP assay was used to confirm the interaction between SOCS2 and miR-1264. (J) SOCS2-AS1 knockdown promoted miR-1264 level. (K) miR-1264 mimics inhibited SOCS2 expression. (L) SOCS2-AS1 knockdown inhibited SOCS2 expression through miR-1264. *P<0.05.