Fig. 2

MCCC2 deficiency induced mitochondrial fusion in CRC cells. A oeMCCC2, KD, and KO cells with respective control staining by the MitoTracker probe were determined to reflect mitochondrial quantity by flow cytometry. MitoTracker probe staining revealed that MCCC2 deficiency led to a decreased number of mitochondria. B Relative mRNA levels of MFN1, MFN2, and OPA1 were quantified using RT-qPCR. MCCC2 deficiency led to the upregulation of mitochondrial fusion markers, whereas MCCC2 overexpression downregulated the expression of these markers. C Western blotting confirmed the protein levels of MFN1, MFN2, and OPA1 in oeMCCC2, KD, and KO cells and their controls, respectively. α-Tubulin and GAPDH served as loading controls. Numbers in the box indicate the relative levels in pairwise semiquantification. D The mitochondria of oeMCCC2, KD, or KO cells, and control cells, were observed by TEM. The images of mitochondria were taken at 15,000× magnification, and a portion of them was enlarged on the right. MCCC2 deficiency promotes mitochondrial fusion, whileMCCC2 overexpression has no apparent effect on mitochondrial morphology. Scale bar: 2 μm. E ATP contents of oeMCCC2, KD, KO cells, and their controls were measured using a luminometric assay and compared in a pairwise fashion. F The GSEA analysis of the transcriptomes (oeMCCC2 versus oeControl cells) showed a significant association between oeMCCC2 and hallmark of glycolysis signaling pathway. NES, normalized enrichment score; FDR, false discovery rate. G The GSEA analysis of the transcriptomes (siMCCC2 versus siControl cells) showed a significant association between siControl and the hallmark of the glycolysis signaling pathway. All data presented as mean ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001