Additional file 2: Figure S1. of Circular RNA circ-ITCH inhibits bladder cancer progression by sponging miR-17/miR-224 and regulating p21, PTEN expression

a Northern blot with 5 mg of RNA from EJ cells transfected with empty vector (GFP) or circ-ITCH vector (circ-ITCH). The blot was probed against circ-ITCH and 18S ribosomal RNA(loading control). b. The overexpression of cir-ITCH had no obvious effect on the expression of its parental gene ITCH using qRT-PCR in BCa cells. c. CCK-8 assay showed that knocking down p21 promoted the proliferation ability of BCa cell EJ. d. CCK-8 assay showed that knocking down PTEN promoted the proliferation ability of BCa cell EJ. (*P < 0.05, Student’s t-test). (ZIP 339 kb)

a. 采用转染空载体（GFP）或circ-ITCH过表达载体的EJ细胞提取的5 mg RNA进行Northern印迹杂交（Northern blot），以circ-ITCH及18S核糖体RNA为探针完成杂交，其中18S核糖体RNA作为上样内参。b. 于膀胱癌细胞（BCa）中采用实时定量逆转录聚合酶链反应（qRT-PCR）检测发现，circ-ITCH过表达对其亲本基因ITCH的表达无显著影响。c. CCK-8细胞增殖实验结果显示，敲低p21可增强膀胱癌细胞EJ的增殖能力。d. CCK-8细胞增殖实验结果显示，敲低磷酸酶和张力蛋白同源物（PTEN）可促进膀胱癌细胞EJ的增殖能力（*P < 0.05，采用学生t检验）。（ZIP格式压缩包，大小339 KB）