Figure 3 from Tumor Vessel Normalization via PFKFB3 Inhibition Alleviates Hypoxia and Increases Tumor Necrosis in Rectal Cancer upon Radiotherapy

Gene set enrichment analysis of 3PO-treated colon organoids. A, Representative pictures of patient-derived tumor organoids treated with 30 µmol/L 3PO for 24 hours. Scale bars, 200 μm. B, Patient-derived tumor organoid size quantification. Data are displayed as means ± SD. ∗, P < 0.05; one-way ANOVA. C, GSEA with gene sets derived from the GO Biological Process ontology. For the enrichment plot (score curves), refer to Supplementary Fig. S2. FDR, false discovery rate. D and E, Protein expression levels of NDUFB6, ACAD9, and LDHA in colorectal cancer organoids (PT-73, PT-70, and PT-79) and colorectal cancer cell lines treated with 3PO for 24 hours. ACTIN and HSC70 were used as loading control. Uncropped Western blot pictures at Supplementary Fig. S5. In all cases, densitometry analysis is performed using loading control as references. F, Seahorse analysis for extracellular acidification rate performed with patient-derived tumor organoids treated with 10 µmol/L 3PO for 24 hours. Data is displayed as SD. ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001.