Fig. 5: Voltage-dependent anion channel 2 (VDAC2) coordinates Bid phosphorylation-dependent apoptotic priming in mitosis. | Cell Death & Disease

Fig. 5: Voltage-dependent anion channel 2 (VDAC2) coordinates Bid phosphorylation-dependent apoptotic priming in mitosis.

From: BioID-based proteomic analysis of the Bid interactome identifies novel proteins involved in cell-cycle-dependent apoptotic priming

Fig. 5

A Single MCF-7 cell clones were grown following CRISPR/Cas9 targeting of VDAC2. Left-hand panel—agarose gel electrophoresis of 300-bp PCR product at VDAC2 CRISPR target PAM site. Right-hand panel—table displaying indels caused by CRISPR/Cas9 determined by direct DNA sequencing of the amplified regions. B Single-cell-fate profiles of the three MCF-7 VDAC2 KO lines in A, untreated (control) or treated with 1 μM Taxol over 48 h. Data represent 90 cells over three independent experiments. Percentage of death in mitosis is shown for Taxol-treated cells. C D11 VDAC2 KO MCF-7 cell lines were generated stably expressing VDAC2-V5, shBid alone or shBid in conjunction with the indicated full-length mouse Bid-GFP variants (mBidWT-GFP, mBidS66A-GFP and mBidG94E-GFP). Cells were either untreated or enriched for mitosis with 18-h treatment in nocodazole followed by shake off. Whole-cell lysates were immunoblotted with the indicated antibodies. β-actin serves as loading control, while phospho-histone H3 (pH 3) indicates mitosis. D Summary of apoptosis in mitosis for single-cell-fate analysis of the cell lines in C, untreated or treated with 1 μM Taxol and/or 5 μM ABT737 over 48 h (single-cell traces in Supplementary Fig. S5C). Data represent 90 cells tracked over three independent experiments. Error bars represent SD. Data analysed by one-way ANOVA, followed by Tukey’s multiple-comparison test. ns = non-significant, **P < 0.01, ***P < 0.001.

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