Fig. 3

Phosphorylations of CDK1 and STAT3 were reduced by fisetin. a Phospho-array analysis of PANC-1 cells treated with fisetin. *Blue and red squares represented phosphorylation of STAT3-Y705 and CDK1-Y15, respectively. b KEGG pathway enrichment of proteins with differential levels of phosphorylation after fisetin treatment. Red asterisk (*) indicated that CSC related pathways as HIF-1 signaling and JAK–STAT signaling were enriched by KEGG pathway analysis. c, d Heat map representing phosphorylation of proteins enriched in HIF-1 signaling and JAK–STAT signaling pathways (data are presented as mean, n = 2). e, f Integrated transcriptome and phospho-array analysis of HIF-1 signaling and JAK–STAT signaling pathways in PANC-1 cells treated with fisetin. Red, green and blue spots indicated that both mRNA and phosphorylation of proteins decreased (mRNA: FC ≥ 2, phosphorylation: FC ≥ 1.12, P < 0.05), only phosphorylation of proteins reduced, mRNA increased and phosphorylation of proteins decreased, respectively. g Western blot analysis was used to test the phosphorylation of CDK1 and STAT3 in PDAC cells treated by fisetin. h Expressions of p-CDK1 (Y15) and p-STAT3 (Y705) in control and fisetin treatemt groups were examined by immunofluorescence. Scale Bars 15 μm. i, j The survival analysis of patients with PDAC from GEPIA database. STAT3 is highly expressed in pancreatic cancer tissue. High levels of STAT3 are associated with poor outcomes. Data are presented as mean ± SD (n = 89). T, tumor; N, tumor-adjacent tissue. *P < 0.05