Fig. 6: VDAC1 was associated with fatty acid synthesis-related mtDNA release and EC dysfunction during sepsis.

A, B HUVEC were pretreated with C75 before LPS stimuli. The levels of VDAC1 oligomerization were detected by western blot (n = 3). C, D VDAC1 were knocked down with siRNA before LPS stimuli. The levels of VDAC1 oligomerization were detected by western blot (n = 3). E, F Immunoblot showing the expression levels of VCAM-1, E-selectin, PAI-1 and TF in HUVEC pretreated with VBIT-4 (40 µM) before LPS stimuli (n = 3). G, H HUVEC were stimulated with LPS in the presence or absence of VBIT-4 (10 µM, 40 µM). The protein levels of VDAC1 oligomerization were analyzed by western blot (n = 3). I VDAC1 in HUVEC was knocked down with siRNA before LPS stimuli for 4 h. Cytosolic mtDNA/nDNA was detected by RT-qPCR (n = 5). J, K VDAC1 was knocked down before LPS stimuli. Confocal laser microscope assay was used to assess the levels of mtDNA (anti-dsDNA, green). Mitochondria were stained in red with Mito-Tracker and nuclei were stained in blue with DAPI. Quantitative results from 8 cells for each condition were reported. L HUVEC were stimulated with LPS in the presence or absence of VBIT-4 (40 µM). Cytosolic mtDNA/nDNA was detected by RT-qPCR (n = 5). M, N HUVEC were pretreated with VBIT-4 (40 µM) before LPS stimuli. Confocal laser microscope assay was used to assess the levels of mtDNA (anti-dsDNA, green). Mitochondria were stained in red with Mito-Tracker and nuclei were stained in blue with DAPI. Quantitative results from 8 cells for each condition were reported. All data were expressed as the mean ± SD. Comparison among three or more groups was analyzed by one-way ANOVA. ns no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.