Fig. 4. PFKFB3 mediates tumour vessel normalisation by promoting CXCL8 transcription and secretion in HER2⁺ gastric cancer.

a Gene-set enrichment analysis (GSEA) of the protein profiles in between PFKFB3^high tumours and PFKFB3^low tumours with normalised P values ≤ 0.05, false discovery rate q values ≤ 0.1, and normalized enrichment scores ≥ 1. b Analysis of proteome profiler antibody in culture supernatant of WTs and TRs. Hotspots of proteins are highlighted. c, d RT-PCR and Western blot were used to evaluate the function of PFKFB3 in regulating CXCL8 expression. e The secretion level of CXCL8 was analysed by ELISA assays between sensitive and resistant cells. f Representative images of PFKFB3 and CXCL8 IHC staining of PFKFB3^high tumours and PFKFB3^low tumours (Left). Association between PFKFB3 and CXCL8 expression score (Right). g–i RT-PCR and tube-formation analysis to evaluate vessel branching, maturation and quiescence in EC cocultured with (NC vs. shPFKFB3), (Vector vs. PFKFB3) and the function of CXCL8 recombination protein and inhibitor SB225002. Values are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, based on Student’s t test.