Additional file 1 of RNA demethylase ALKBH5 prevents pancreatic cancer progression by posttranscriptional activation of PER1 in an m6A-YTHDF2-dependent manner
2020-05-20T03:50:23Z (GMT) by
Additional file 1: Figure S1. Construction of the ALKBH5-manipulated pancreatic cancer (PC) cell lines. (a) The mRNA expression of ALKBH5 in HPDE6c7 and five PC cell lines. (b) Protein level of ALKBH5 in HPDE6c7 and five PC cell lines. (c) Examination of a stable overexpression of ALKBH5 in BxPC-3 cells (left panel) and stable knockdown of ALKBH5 in SW1990 cells (right panel). **P < 0.01, ***P < 0.001 (t test compared with HPDE6c7 group). Figure S2. m6A level in BxPC-3/Vector and BxPC-3/Lv-ALKBH5 cells. Overexpression of ALKBH5 significantly reduced m6A level in BxPC-3 cells.*P < 0.05. Figure S3. The semi-quantitative analysis of western blotting results in Fig. 3. (a) ALKBH5 overexpression leads to the upregulated protein level of p-CDK1 and downregulated protein level of Cyclin B1 in BxPC-3cells. (b) The protein level of p-CDK1 and Cyclin B1 was significantly downregulated and upregulated according to the knocking down of ALKBH5 in BxPC-3cells. ***P < 0.001. Figure S4. The semi-quantitative analysis of western blotting results in Fig. 7i. ALKBH5 knockdown resulted in the downregulation of genes promoting G2/M phase cell cycle arrest and up-regulation of CYCLIN B1 on a basis of PER1–loss, whereas PER1 overexpression partially reversed these abnormalities. *P < 0.05, **P < 0.01, ***P < 0.001. Table S1. Primers used in this study. Table S2. 367 differentially expressed genes.