Journal: American Journal of Translational Research

Article Title: S-phase kinase-associated protein 2 impairs the inhibitory effects of miR-1236-3p on bladder tumors

doi:

Figure Lengend Snippet: Primers used in this study

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) (Sigma-Aldrich, USA) and then incubated with primary antibodies, including those against Skp2 (1/1000) (Affinity, USA), P21 (1/1000) (BD Biosciences), P27 (1/1000) (Affinity, USA), CDK1 (1/1000) (Boster, China), GAPDH (1/500) (Boster, China) and β-actin (1/500) (Boster, China), overnight at 4°C.

Techniques: Control

Journal: American Journal of Translational Research

Article Title: S-phase kinase-associated protein 2 impairs the inhibitory effects of miR-1236-3p on bladder tumors

doi:

Figure Lengend Snippet: miR-1236 induced Skp2 expression independent of p21 activation and influenced downstream Skp2 gene expression. A. Skp2 mRNA levels were evaluated by qRT-PCR. **P<0.01 compared to control miRNA group. B. Expression of Skp2 protein was further detected by Western blot analysis. Only miR-1236 up-regulated Skp2 expression. C. Expression of p21 and Skp2 mRNA in BCa cells was detected by qRT-PCR. *P<0.05, **P<0.01 compared to control miRNA group. D. Western blotting was conducted to detect the expression of p21 and Skp2 protein in BCa cells. Knockdown with sip21 had no influence on the up-regulation of Skp2 by miR-1236. E. mRNA expression levels of p27 and CDK1 were assessed by qRT-PCR. *P<0.05 compared to the control miRNA group. F. P27 and CDK1 expression levels were detected by Western blot in 5637 and T24 cells. miR-1236 decreased p27 expression and up-regulated CDK1 expression. dsRNA-245 had no significant effect on p27 and CDK1. G. Expression of Skp2, p27 and CDK1 was detected by qRT-PCR. GAPDH served as an internal control. **P<0.01, ***P<0.001 compared with the miR-1236 group. H. Skp2, p27 and CDK1 protein expression levels were detected by Western blot. β-actin levels were used as an internal control. miR-1236 influenced p27 and CDK1 expression through up-regulation of Skp2.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) (Sigma-Aldrich, USA) and then incubated with primary antibodies, including those against Skp2 (1/1000) (Affinity, USA), P21 (1/1000) (BD Biosciences), P27 (1/1000) (Affinity, USA), CDK1 (1/1000) (Boster, China), GAPDH (1/500) (Boster, China) and β-actin (1/500) (Boster, China), overnight at 4°C.

Techniques: Expressing, Activation Assay, Gene Expression, Quantitative RT-PCR, Control, Western Blot, Knockdown