Fig. 2: Mitochondrial dysfunction and lipid metabolism disorder induced by CdtB in hepatocytes. | Nature Communications

Fig. 2: Mitochondrial dysfunction and lipid metabolism disorder induced by CdtB in hepatocytes.

From: Helicobacter hepaticus promotes hepatic steatosis through CdtB-induced mitochondrial stress and lipid metabolism reprogramming

Fig. 2

a Immunofluorescence images of mitochondrial morphology (TOM20, green) in Hep3B cells transfected with pCMV-vector or pCMV-CdtB (CdtB mAb, red) for 24 h. b Transmission electron microscopy images of liver sections from mice with rAd-vector or rAd-CdtB tail vein injection (i.v.) at 7 days post-infection (DPI). Red triangle: Damaged mitochondria with loss of ridges and reduced inner and outer membrane integrity; Yellow triangle: Lipid droplet; Green triangle: Autophagosome; Blue triangle: Autophagosome containing damaged mitochondria; Purple triangle: Autolysosome. c MitoSOX Red staining of mitochondrial superoxides in HEK293A cells transfected with pCMV or pCMV-EGFP-CdtB. d ATP production in Huh-7 cells transfected with pCMV-vector, pCMV-CdtB or pCMV-COX8A-CdtB for 24 h (n = 3 independent experiments). e, f Analysis of mitochondrial DNA (mtDNA) in purified cytoplasmic fractions of Hep3B cells transfected with pCMV or pCMV-CdtB (e), followed by treatment with vehicle control (NC) or CsA (500 ng/mL) (f) for 24 h (n = 3 independent experiments). g Serum mtDNA level following intravenous administration of mice with rAd-vector or rAd-CdtB (n = 5 mice). h Flow cytometry analysis of Hep3B cell death labeled with propidium iodide (PI) following various treatments. i Flow cytometry analysis of mitochondrial membrane potential by tetramethylrhodamine methyl ester (TMRM) in Hep3B cells (n = 3 independent experiments) transfected with pCMV-EGFP or pCMV-EGFP-CdtB or treated with cis-Platinum (CDDP) (10 μM) for 24 h. j Western blot analysis of CPT1a expression of Hep3B cells infected with wild type (WT), or CdtB knockout (ΔCdtB) H. hepaticus. Scale bars represent 2 μm (b, original) or 1 μm (b, enlarged), 10 µm (a) or 50 µm (c). Statistical analysis was determined by one-way ANOVA (d, i), two-way ANOVA (f, g) followed by Dunnett’s multiple comparisons test, or unpaired two-tailed Student’s t test (e). Source data are provided as a Source Data file.

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