Figure EV1. p13 knockdown prevents mitochondrial dysfunction and cytotoxicity.

• A, B
  Decreased expression of p13 mRNA (A) or p13 protein (B) in cells 72 or 96 h after infection with lentiviral vectors expressing p13 shRNA. mRNA levels were quantified by real‐time RT–PCR (A). The levels of cleaved PARP were normalized to those of β‐actin (B). Representative images (left) and their quantification (right) were shown.
• C
  Decreased expression of p13 mRNA in cells 96 h after infection with lentiviral vectors expressing p13 shRNA #2. mRNA levels were quantified by real‐time RT–PCR.
• D–F
  Prevention of the rotenone‐induced decrease in ΔΨm (D), increase in PARP cleavage (E) and decrease in complex I activity (F) in cells infected with lentiviral vectors expressing p13 shRNA #2. Complex I activity was measured on the basis of NADH‐oxidizing activity. Seventy‐two hours after infection, cells were exposed to vehicle or 100 nM rotenone for 24 h.
• G, H
  Prevention of the MPP⁺‐induced decrease in ΔΨm (G) and release of LDH (H) in p13 shRNA‐infected cells. LDH release was measured in culture medium of SH‐SY5Y cells using the LDH assay kit. Seventy‐two hours after infection, cells were treated with vehicle or 5 mM MPP⁺ for 24 h.
• I
  Decreased expression of PINK1 mRNA in cells 96 h after infection with lentiviral vectors expressing PINK1 shRNA. mRNA levels were quantified by real‐time RT–PCR.
• J, K
  Prevention of the PINK1‐knockdown‐induced decrease in ΔΨm (J) and increase in PARP cleavage (K) in p13 shRNA‐infected cells 96 h after infection with lentiviral vectors.

Data information: ΔΨm was determined by measuring the TMRE fluorescence levels (D, G, J). The levels of cleaved PARP were normalized to those of β‐actin (E, K). Representative images (left) and their quantification (right) were shown. All data are presented as the mean ± SEM (n = 3). *P < 0.05, **P < 0.01 by Student's t‐test (A–C and I) or the Tukey–Kramer test (D–H, J and K).