Fig. 4

lnc-RP11-536 K7.3 recruit SOX2 to regulate the promoter activity of USP7. A GSEA was performed in lnc-RP11-536 K7.3-KO organoids and control group. The gene signature was defined by genes with significant expression changes. B qRT-PCR assay of USP7 mRNA in lnc-RP11-536 K7.3-KO organoids and cells and control groups (^**P < 0.01). The experiment was repeated 3 times. C Luciferase reporter assay indicated that USP7 activity was affected by knockout of lnc-RP11-536 K7.3 (^**P < 0.01). The experiment was repeated 3 times. D RNA pull-down assay followed by silver staining and western blot was performed on chemo-resistant CC organoids. E qRT-PCR was carried out after completion of RNA RIP assay. The experiment was repeated 3 times. F FISH and immunofluorescence assay detected the expressions of lnc-RP11-536 K7.3 and SOX2 in lnc-RP11-536 K7.3 knocked out organoids and CR-RKO cell lines and controls. The experiment was repeated 3 times. G-H The map of SOX2 binding sits in the promoter region of USP7 and the results of ChIP analysis showed that SOX2 can bind to the USP7 promoter region. I-K Luciferase reporter assay of SOX2 mutant sites in the promoter region of USP7. (^**P < 0.01). The experiment was repeated 3 times. L qRT-PCR assay of USP7 mRNA in knockout of lnc-RP11-536 K7.3 and overexpression of SOX2 organoids and cells and control groups (^**P < 0.01). The experiment was repeated 3 times. M CHIP results showed that SOX2 could bind to USP7 and mutation of SOX2 binding motifs abrogated its transcriptional regulation on USP7. N Luciferase reporter assay indicated that USP7 activity was affected by knockout of lnc-RP11-536 K7.3 and overexpression of SOX2 (^**P < 0.01). The experiment was repeated 3 times