Fig 1: Nuclear PKM2 interacts with FOSL1 to establish super-enhancers and promotes VM in colorectal cancer. A, Proliferation of PKM2 WT–overexpressing HCT116 and SW620 cells with FOSL1 knockdown as indicated was analyzed using CCK8 assay. B, Statistical analysis of the colony formation assay in Supplementary Fig. S6B. All of the experiments were performed in triplicate, and data are shown as means ± SD. C, Statistical analysis of the soft-agar colony formation assay in Supplementary Fig. S6C. All of the experiments were performed in triplicate, and data are shown as means ± SD. D, Statistical analysis of the tube formation assay in Supplementary Fig. S6D. All of the experiments were performed in triplicate, and data are shown as means ± SD. E, Photograph and comparison of tumor sizes in different groups. Bevacizumab, 5 mg/kg every 3 days. F, Growth curves of tumors formed by indicated cells. G, Tumor weights in the indicated groups. H, Representative IHC staining for CD31, PAS, and Ki-67 in mouse tumors. Scale bar, black, 50 μm. I, Quantification of the number of VM channels (left) and proliferation index (right) in the indicated xenograft tumors; proliferation index was determined using the percentage of Ki-67–positive cells. J, CUT&Tag data showed co-occupancy for PKM2, FOSL1, and H3K27ac across the genome in HCT116 cells. K, Heatmap showing the reduced signal intensity of H3K27ac of FOSL1 targets in PKM2 K206R compared with PKM2 WT HCT116 cells. L, Genomic distribution of H3K27ac sites in PKM2 WT or K206R HCT116 cells. M, Genomic distribution of FOSL1 and PKM2 sites. N, Co-localization peaks of FOSL1 and PKM2 at genome-wide. O, Genome browser tracks of CUT&Tag data for H3K27ac, FOSL1, and PKM2 and CUT&Tag and ATAC-seq data for PKM2 WT and PKM2 K206R H3K27ac at SE regions of FOSL1 (left) and SNAI2 (right) in HCT116 cells. P, ChIP-qPCR analysis for FOSL1 status in the SE regions of FOSL1 and SNAI2 of PKM2 WT or K206R HCT116 cells treated with 5 mmol/L Nala for 24 hours as indicated. Q, Expression of FOSL1, SNAI2, MAZ, and MYC was detected in PKM2 WT or K206R HCT116 cells treated with 5 mmol/L Nala for 24 hours as indicated by RT-qPCR. ns, no significance; **, P ≤ 0.01; ***, P ≤ 0.001. UTR, untranslated region.