Fig. 4

Identification of the SGMS1-AS1/miR-181d-5p/SRPK2 ceRNA network activated by BP1 overexpression. a Prediction of the location of SGMS1-AS1 by IncLocator (http://www.csbio.sjtu.edu.cn/bioinf/lncLocator). b Validation of the location of SGMS1-AS1 by RNA FISH. SGMS1-AS1 was localized mainly in the cytoplasm of K562 cells. c Prediction of the downstream miRNA of SGMS1-AS1 by Venn analysis of LncBase Predicted (http://carolina.imis.athena-innovation.gr/diana_tools/web/index.php?r=lncbasev2%2Findex), starBase (http://starbase.sysu.edu.cn/), and miRNA-Seq. d Validation of the selected gene expression by RT-qPCR. e Dual luciferase experiment of SGMS1-AS1 binding to miR-181d-5p. Overexpression of miR-181d-5p significantly reduced the luciferase activity of the wild-type SGMS1-AS1 vector but not of the mutated SGMS1-AS1 vector. f Prediction of the downstream mRNA of SGMS1-AS1 by Venn analysis of mRNA-Seq, miRDB (http://mirdb.org/miRDB/), TargetScan (http://www.targetscan.org/vert_72/), mirDIP (http://ophid.utoronto.ca/mirDIP/index.jsp), and starBase (http://starbase.sysu.edu.cn/). g Validation of the selected gene expression by RT-qPCR. h Dual luciferase experiment of miR-181d-5p binding to SRPK2. Overexpression of miR-181d-5p markedly reduced the luciferase activity of the wild-type 3′-UTR of SRPK2 vector but not of the mutated 3′-UTR of SRPK2 vector. i The SGMS1-AS1/miR-181d-5p/SRPK2 expression detected by RT-qPCR after Ago2-RIP. Endogenous SGMS1-AS1, miR-181d-5p, and SRPK2 was preferentially enriched in Ago2-RIPs compared with control IgG-RIPs. j RNA immunoprecipitation (RIP) efficiency confirmation detected by western blot. *P < 0.05; **P < 0.01; ***P < 0.001