Fig. 4: Nuclear MYG1 plays a more critical role in promoting CRC progression through glycolysis.

a MYG1 expression in the subcellular compartments of cells were determined by western blot. W, whole cell lysis. C, cytoplasm. N, nucleus. M, mitochondria. CM, cytoplasm with mitochondria removed. The samples derive from the same experiment but different gels for MYG1 and COX4, and another for GAPDH and H3 were processed in parallel. b Mitochondrial fractions were analyzed by protease K shaving assay. VDAC1 was sensitive to protease K treatment, whereas TIMM23 was resistant. Proteins were digested by protease K in the presence of 1% Triton X-100. The samples derive from the same experiment but different gels for MYG1 and VDAC1, and another for TIMM23 were processed in parallel. c Images of Flag-tagged MYG1 in mitochondria of 293 T cells as detected by immunoelectron microscopy. Scale bar, 200 nm (left) and 500 nm (right). Representative of 25 images from n = 2 independent experiments. d Schematic of MYG1 domain and variant constructs. e Cellular location of Flag-tagged MYG1 variants was confirmed by IF in HCT116 cells. Scale bar, 10 μm. Representative images from n = 2 independent experiments. f The levels of glucose uptake and lactate secretion were detected. g–k Luciferase-labeled HCT116 were utilized to establish orthotopic CRC mouse models (n = 5 in each group). Bioluminescence imaging of mice was detected on the 30th day (g) and the signals were quantified (h). PET/CT images with the signals indicated SUV normalized to body weight (SUVbw) of mice. The maximum ¹⁸F-FDG uptake value (SUVmax) was obtained by browsing different layers of PET imaging (i, dashed cycle marked) and quantified (j). Tumors were separated and weighted (k) after sacrificing the mice. The invasion (l) and colony formation ability (m) were detected. n = 3 independent experiments (a–b, f, and l–m). Kruskal–Wallis test, Dunn’s multiple comparisons test (h and k). One-way ANOVA, Tukey’s multiple comparisons test (f and j). Two-way ANOVA, Tukey’s multiple comparisons test (l–m). p value was provided in the figure. Error bars, mean ± SD. See also Supplementary Fig. 5. Source data are provided as a Source Data file.