Fig. 7.

AMBRA1 induce mitophagy in an in vitro model of ischaemia. a Lysates of SH-SY5Y cells exposed to different time points of OGD/RX (0 min, 30 min, 1 h and 3 h) were blotted with the indicated antibodies; n = 3. b Endogenous AMBRA1 levels were downregulated using siRNA-AMBRA1 in SH-SY5Y cells. Following exposure to 3 h OGD/RX, total lysates were subjected to western blot analysis in order to analyse mitochondria clearance, looking at COXII protein levels; n = 3. c PcDNA3 or Myc-AMBRA1^WT-expressing SH-SY5Y cells were transfected with siRNA-Ctr or siRNA-HUWE1 constructs and then immunoblotted for the indicated proteins; n = 3. d SH-SY5Y cells transfected with PcDNA3 or HA-IKKα^K44M constructs in combination with Myc-AMBRA1^WT were subjected to western blot analysis for the indicated antibodies; n = 3. Data represent the mean of three different experiments (±S.D.). *P < 0.05. Statistical analysis was performed using one-way ANOVA. M_r(K) = relative molecular mass expressed in kilodalton