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. 2018 Jul 26;37(17):e99238. doi: 10.15252/embj.201899238

Figure 1. mtDNA is released from mitochondria following MOMP in a BAX/BAK‐dependent manner.

Figure 1

  1. Fixed super‐resolution Airyscan images of U2OS cells immunostained with anti‐TOM20 (green) and anti‐DNA (red) antibodies. Scale bar = 10 μm. Representative images from three independent experiments.
  2. Airyscan images of U2OS cells treated with 10 μM ABT‐737, 1 μM ActD and 20 μM qVD‐OPh for 3 h, immunostained with anti‐TOM20 and anti‐DNA antibodies. Scale bar = 10 μm. Representative images from three independent experiments.
  3. Quantification of cells exhibiting > 10% mtDNA release following treatment with 10 μM ABT‐737, 1 μM ActD and 20 μM qVD‐OPh. Data are expressed as mean ± SD from three independent experiments and analysed by Student's t‐test.
  4. Quantification of the extent of mitochondrial DNA (mtDNA) nucleoid release per cell following treatment with 10 μM ABT‐737, 1 μM ActD and 20 μM qVD‐OPh. Data are expressed as mean ± SD from two independent experiments and analysed by Student's t‐test.
  5. Airyscan images of MEF cells untreated or treated with 10 μM ABT‐737 and 20 μM qVD‐OPh for 3 h, immunostained with anti‐TOM20 and anti‐DNA antibodies. Scale bar = 10 μm. Representative images from three independent experiments.
  6. BAX and BAK expression levels in U2OS cells with CRISPR‐Cas9‐mediated deletion of BAX, BAK or BAX/BAK.
  7. U2OS cells with BAX, BAK or BAX/BAK deletion by CRISPR‐Cas9 treated with 10 μM ABT‐737 and 2 μM S63845 and analysed for cell viability using an IncuCyte live‐cell imager and SYTOX Green exclusion. Data are expressed as mean ± SEM, representative of three independent experiments, and have been normalised to starting confluency.
  8. Airyscan images of U2OS (i) control cells or with CRISPR‐Cas9‐mediated deletion of either (ii) BAX, (iii) BAK or (iv) BAX and BAK treated with 10 μM ABT‐737, 2 μM S63845 and 20 μM qVD‐OPh for 3 h. Scale bar = 10 μm. Representative images from three independent experiments.
  9. Quantification of mtDNA nucleoid release per cell in U2OS EMPTYCRISPR, BAX CRISPR, BAK CRISPR and BAX/BAK CRISPR cells. Data are expressed as mean ± SD from three independent experiments and analysed using Student's t‐test.

Source data are available online for this figure.