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. 2018 Jan 25;19(3):e44860. doi: 10.15252/embr.201744860

Figure 3. p13 knockdown maintains complex I activity in rotenone‐treated SH‐SY5Y cells.

Figure 3

  • A
    Prevention of the rotenone‐induced decrease in complex I activity in p13 shRNA‐infected cells. Complex I activity was measured on the basis of NADH‐oxidizing activity. Data are presented as the mean ± SEM (n = 3). *P < 0.05 by the Tukey–Kramer test.
  • B, C
    Physical interaction between overexpressed p13 (B) or endogenous p13 (C) and complex I proteins. Lysates were immunoprecipitated with anti‐complex I antibody and control IgG. The lysates and immunoprecipitates were subjected to Western blotting with antibodies against FLAG (B), p13 (C) or NDUFB8 (complex I protein). Cells were infected with lentiviral vectors expressing mock or FLAG‐tagged p13 (p13 o/e, B).
  • D
    Prevention of rotenone‐induced impairment of complex I assembly by p13 knockdown in p13 shRNA‐infected cells. Mitochondrial fractions were subjected to blue native PAGE, followed by Western blotting with an antibody against NDUFB8 (complex I protein). Levels of p13 and Hsp60 (for loading control) in the mitochondrial fraction were analysed.
Data information: Seventy‐two hours after infection with lentiviral vectors expressing scrambled shRNA (Scr shRNA) or p13 shRNA, cells were stimulated with vehicle or 100 nM rotenone for 24 h (A, D).