Fig. 5

circ_0005529 negatively regulates miR-527 through upregulating Sp1. a The intersection of potential genes that could be targeted by miR-527, as predicted by Targetscan, mirTarBase and miRwalk. b Relative luciferase activities of the 3′-UTR reporter plasmids (SLC12A7, KIAA0513, KLHL15, FZD5, UBE2A, LPP, MECP2, ZBTB38, RAB14, PRUNE2, CNEP1R1, Sp1, SH3BP4, HOOK3, H3F3B, PEX26, PFN2, H3F3C, ZMAT3, ATF7IP, ZNF618, MLLT6, PAFAH1B2 and HNRNPA3) were measured in MKN-45 cells upon transfection of miR-527 mimics or NC mimics. c Seeds match for miR-527 in the 3′-UTR of Sp1. The predicted seed-recognition sites in the Sp1 mRNA sequence and the corresponding miR-527 sequence are marked in green. d Relative luciferase activity of the Sp1 3′-UTR reporter plasmid was measured in HGC-27 cells after expressing NC inh or miR-527 inh. The mutant Sp1 3′-UTR reporter was used as a negative control. e Relative Sp1 mRNA expression levels in GC tissues (tumor, n = 63) and normal adjacent tissues (adjacent, n = 63), as determined using qRT-PCR. f Pearson’s correlation analysis of the relative expressions between circ_0005529 and Sp1 mRNA, miR-527 and Sp1 mRNA in GC patients. g Protein expression levels of Sp1 in HGC-27 cells (Lv-NC+miR-NC, Lv-circ_0005529+miR-NC, Lv-circ_0005529+miR-527) and in MKN-45 cells (sh-NC+NC inh, sh-circ_0005529+NC inh, sh-circ_0005529+miR-527 inh), as determined using western blotting. Bands were quantified and shown in histogram. (Mean ± SEM, * p < 0.05, ** p < 0.01)