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. 2017 Jun 12;9(8):1030–1051. doi: 10.15252/emmm.201607088

Figure 4. Mitochondrial ceramide is involved in HPV‐E7‐mediated lethal mitophagy.

Figure 4

  • A, B
    Effects of siRNA‐mediated knockdown of HPV‐E6/E7 on mitophagy in response to C18‐pyr‐cer (20 μM, 1 h) were measured by live cell imaging confocal micrographs of UM‐SCC‐47 cells stained with LTG and MTR (A) or by mitochondrial respiration (oxygen consumption rate) using Seahorse analyzer (B). Scr‐siRNA‐transfected and/or vehicle‐treated cells were used as controls. Data are means ± SD from three independent experiments, analyzed by unpaired Student's t‐test (n = 3, *P = 0.021, **P = 0.0004). In (A), images were quantified using ImageJ, and scale bars represent 100 μm.
  • C–E
    Roles of ectopic expression of HPV‐E6 or E7 in UM‐SCC‐22A cells for enhancing mitochondrial C18‐pyr‐cer‐mediated cell death and mitophagy were measured by trypan blue exclusion assay (C) and Seahorse analyzer for measuring mitochondrial respiration (oxygen consumption rate) (D) or using live cell imaging confocal micrographs of UM‐SCC‐22A cells stained with LTG and MTR (E). Vector‐transfected and/or vehicle‐treated cells were used as controls. Data are means ± SD from three independent experiments, analyzed by unpaired Student's t‐test (n = 3, *P = 0.0093, **P = 0.0072, ***P = 0.00026). In (E), images were quantified using ImageJ, and scale bars represent 100 μm.