Fig. 1: VZV infection results in mitochondrial fission and activates PINK1/Parkin-dependent mitophagy. | Cell Death & Disease

Fig. 1: VZV infection results in mitochondrial fission and activates PINK1/Parkin-dependent mitophagy.

From: Varicella zoster virus glycoprotein E facilitates PINK1/Parkin-mediated mitophagy to evade STING and MAVS-mediated antiviral innate immunity

Fig. 1

A MRC5 cells were infected with mock (m) or VZV YC01 (MOI 0.001). Representative transmission electron microscope images of mock or VZV-infected cells. The red arrow indicates mitochondria. Scale bar = 1 μm. B Quantitative analysis of mitochondria lengths in mock (m) vs. VZV-infected MRC5 cells. The length of ten mitochondria per cell present in more than five cells in each group was measured. Statistical analysis, *p < 0.05 vs. mock-infected cells. C MRC5 and HaCaT cells were infected with mock (m) or VZV (MOI 0.001) for indicated times. Relative tetramethylrhodamine methyl ester (TMRM) intensity was measured (mean ± SD; n = 3). *p < 0.05; **p < 0.01; ***p < 0.001 vs. mock-infected cells. D Immunoblot analysis of VZV gE, p62, LC3, and TOM20 expression in VZV (MOI 0.001)-infected cells at the indicated time points. β-actin was used as a protein loading control. Numbers below the blot represent quantified band intensity by densitometric analysis. E MRC5 and HaCaT cells were infected with mock (m) or VZV (MOI 0.001) and subsequently stained with MitoTracker (green) and LysoTracker (red). % cells with LysoTracker and MitoTracker colocalization were calculated and are presented in the graph. **p < 0.01; ***p < 0.001 vs. mock-infected cells. F Representative confocal analysis of VZV-infected HeLa-Parkin cells expressing mtKeima. Cells were treated with 25 μM CCCP for 2 h or infected with VZV for 48 h. Representative mtKeima fluorescence images are shown and the graph on the right demonstrates mitophagy index. ***p < 0.001 vs. mock-infected control cells (Ctl). G Representative confocal images of HaCaT cells transfected with siCtl or siPINK1, followed by VZV infection (MOI 0.001). The graph shows the percentage of cells displaying mitolysosomes by assessing LysoTracker and MitoTracker colocalization. A minimum of one hundred cells per condition were counted in three independent experiments. **p < 0.01 vs. siCtl-expressing mock-infected cells. H HaCaT cells were transfected with control (siCtl) or Parkin siRNA (siParkin), followed by empty vector (EV) or Parkin-MYC plasmids (siParkin+Parkin) transfection. After transfection, cells were infected with VZV (MOI 0.001) and stained with LysoTracker and MitoTracker to examine the formation of mitolysosomes at 48 hpi. *p < 0.05; **p < 0.01 vs. siCtl-expressing cells, ##p < 0.01 vs. siParkin-expressing EV-transfected cells.

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