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. 2014 Nov 28;16(1):97–106. doi: 10.15252/embr.201438976

Figure 3.

Figure 3

YME1L degradation is initiated by reductions in intracellular ATP and OMA1 activation

  1. Top, bar graph showing the intracellular ATP levels in HEK293T cells incubated with 2-deoxyglucose (2-DG; 10 mM) and/or CCCP (50 μM) for 6 h, as indicated. Error bars show SEM for n = 3. Middle, representative immunoblot of lysates prepared from HEK293T cells incubated with 2-deoxyglucose (2-DG; 10 mM) and/or CCCP (50 μM) for 6 h, as indicated. The long and short isoforms of OPA1 are indicated by the arrows. Bottom, quantification of YME1L (red) and AFG3L2 (blue) from immunoblots as shown above. Error bars show SEM for n = 3. *P-value < 0.05.
  2. Top, bar graph showing the intracellular ATP levels in HEK293T cells cultured in galactose-supplemented media incubated with oligomycin A (OA; 5 nM) and/or CCCP (50 μM) for 6 h, as indicated. Error bars show SEM for n = 3. Middle, representative immunoblot of lysates prepared from HEK293T cells cultured in galactose-supplemented media incubated with oligomycin A (OA; 5 nM) and/or CCCP (50 μM) for 6 h, as indicated. The long and short isoforms of OPA1 are indicated by the arrows. Bottom, quantification of YME1L (red) and AFG3L2 (blue) from immunoblots as shown above. Error bars show SEM for n = 3. *P-value < 0.05.
  3. Immunoblot of lysates from HEK293T cells incubated with 2-deoxyglucose (2-DG; 10 mM) and/or CCCP (50 μM) for the indicated times. Full-length OMA1 (FL) and a cleaved, proteolytically active OMA1 fragment are shown by the arrows.
  4. Immunoblot of lysates from SHSY5Y cells treated with H2O2 (200 μM) for the indicated times.
  5. Illustration depicting the predicted mechanism for stress-induced YME1L degradation.

Source data are available online for this figure.