Fig. 5

Hsp70-Bim PPI induced mitophagy independent on Bax and Bak. A Western blot analysis of LC3 level in isolated mitochondria from MEFs and Bax/Bak DKO MEFs treated with HBSS in the presence or absence of 10 μM S1g-2 for 4 h. An equivalent of DMSO were added to the compound untreated group as vehicle control. **P < 0.01 (one-way ANOVA test). B Bax/Bak DKO MEFs transfected with Bim or the respective vector control was treated with 5 mg/mL and 10 mg/mL dox respectively for 48 h, and then the cell lysates were applied for Western blot analysis of Bim. The harvested cells were applied for mitochondria isolation and the LC3 level was analyzed by Western blotting. **P < 0.01 (one-way ANOVA test). C Co-IP analysis of Hsp70 interactions with Bim, TOMM20 and parkin in Bax/Bak DKO MEFs treated as described in B. **P < 0.01 (one-way ANOVA test). D Co-IP analysis of TOMM20 interactions with parkin and TOMM20 ubiquitination which was visualized by Western blot analysis using anti-ubiquitin in Bax/Bak DKO MEFs treated as described in B. All figures represent the results from n = 3 biologically independent experiments. **P < 0.01 (one-way ANOVA test)