Table 3.

Densitometry of VDAC1 oligomers identified by BN-SDS PAGE following Bax 1D1 immunodepletion.

Oligomer	VDAC: Healthy neurons		VDAC: Apoptotic neurons
	control deplete	1D1 deplete	control deplete	1D1 deplete
1	5.37±1.29	7.73±1.08	5.39±2.14	4.55±1.83
2	0.66±0.08	0.95±0.26	0.61±0.32	0.37±0.13
3	0.59±0.10	0.74±0.29	0.56±0.23	0.51±0.10
4	0.94±0.17	1.06±0.35	0.99±0.16	2.30±1.27
5	1.84±0.18	3.18±1.14	2.82±0.84	4.54±2.37

Digitonin solubilized extracts prepared from CGNs exposed 5h to high (healthy) or low (apoptotic) K were incubated overnight at 4°C with a Bax 1D1 or control mouse monoclonal antibody. The extracts were cleared of antibody complexes using Protein G Dynabeads then subjected to BN-SDS PAGE and western blotting as indicated in Fig. 4. VDAC1 oligomer intensities after immunodepletion were calculated using Image J software as the sum of pixel intensities (in optical density units) within the elliptical area of each spot. Statistical analysis by 2-way ANOVA indicated no significant effects of antibody [control, 1D1] (p= 0.77, 0.89, 0.83, 0.24, and 0.25 for oligomers 1, 2, 3, 4, and 5, respectively), treatment [high, low K] (p= 0.40, 0.18, 0.56, 0.29, and 0.37 for oligomers 1, 2, 3, 4, and 5, respectively), or antibody-treatment interaction (p= 0.41, 0.25, 0.64, 0.33, and 0.88 for oligomers 1, 2, 3, 4, and 5, respectively) on the abundance of the VDAC1 oligomers. Bax oligomer depletion therefore does not significantly affect the amount of each corresponding VDAC1 oligomer. Data are mean ± SEM of three (low K) or four (high K) experiments.