Fig. 4

CircSTX6 serves as a miR-515-3p sponge to upregulate SUZ12 expression. A Heat map showed the differentially expressed genes in EJ and UMUC3 cells after knockdown of circSTX6. B Venn diagram showed the genes detected by RNA-seq and overlapping analysis with the target mRNA of miR-515-3p predicted by miRTarBase, miRDB and TargetScan databases. C Schematic of SUZ12 3’ UTR wild-type (wt) and mutant (mut) luciferase reporter vectors. D The relative luciferase activities were analyzed in BCa cells co-transfected with miR-515-3p or miR-NC and luciferase reporter vectors SUZ12 3’UTR (WT) or SUZ12 3’UTR (Mut). E-F The mRNA and protein level of SUZ12 in EJ cells stably transfected scramble, sh-circ#1, sh-circ#2, vector or circSTX6. G qRT-PCR analysis revealed the levels of SUZ12 in our 16 paired samples of BCa and NAT. N noncancerous adjacent tissues, T tumorous tissue. P values are calculated by paired two-sided t-test. H The expression of SUZ12 was analyzed using western blot in our clinical samples. I The correlation between the transcript levels of circSTX6 and SUZ12 in BCa tissues was analyzed (n = 16). J-K qRT-PCR and western blot were performed to evaluate the expression of SUZ12 in EJ and UMUC3 cells which were transfected with scramble or sh-circ#1, and those co-transfected with anti-miR-NC or anti miR-515-3p. The data are presented as the means ± S.D. of at least three independent experiments. *P < 0.05. P values are calculated by Student’s t test in F and one-way ANOVA in D, E and J