Fig. 2

CDK1 and cancer stemness related genes were inhibited by fisetin. a Flow chart of nude mice model of CDK1 silencing PANC-1 cells. CDK1-KO: CDK1-knockout. b Photograph and weights of xenograft tumors of wild type (WT) and CDK1-Knockout (KO) mice at day 30 after tumor cells injection. Data are presented as mean ± SD (n = 5); *P < 0.05. c Three-dimensional illustration of docked CDK1–fisetin complex. CDK1 is represented as slate cartoon. Fisetin is showed as cyan sticks, binding sites are displayed a series of pink sticks and hydrogen bonds are represented as yellow dashes. d RMSD plot of CDK1-Fisetin. Cα-RMSD, ligand RMSD of ligand fit on protein or ligand fit on ligand are represented as deep blue, deep red and pink lines, respectively. e Western blot analysis showed that expression of CDK1 were significantly inhibited by fisetin in both pancreatic cancer PANC-1 and HPC-Y5 cells. f Heat map representing mRNA expression levels of stemness related genes in RNA-sequencing of PANC-1 cells. Each column indicates one sample and each row represents one mRNA. Orange and blue indicate high and low expression, respectively. g Western blot analysis was used to determine the protein expressions of Nanog, Oct4 and Sox2 with treatment of fisetin for 48 h. h, i Sphere formation assay of PDAC cells. After 100 μM fisetin treatment for 48 h, PANC-1 and HPC-Y5 cells were subjected to the tumor sphere-formation assay in ultra-low cluster plates. Scale bars, 100 μm. Data are presented as mean ± SD (n = 3),*P < 0.05, **P < 0.01. j Statistical plot of ratio of CD44 + /CD24 + positive cells in control or fisetin treatment PANC-1 cell line. Data are presented as mean ± SD (n = 3); *P < 0.05