Fig 2: Voltage-dependent anion channel 1 (VDAC1) regulation after retinal mechanical injury.Western blot analysis of VDAC1 protein levels A 2 h, B 6 h, C 24 h, 3 days (D), and 7 days (E) after the lesion. Ribosomal L26 protein was used as an internal control. Representative VDAC1 immunofluorescence (IF, green) in transverse sections of F controls, G 2 h, and H 6 h after lesion, counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue). The arrows indicate the epicenter of the lesion, whereas head arrows indicate the accumulation of VDAC1 after the lesion. I Left: validation of cellular fractioning analyzing COX IV, ERp44, and ZO-1 proteins in mitochondria (Mit), plasma membrane (PM) and endoplasmic reticulum (ER). Right: representation of cell organelles according to VDAC1 levels after the mechanical trauma. J Crosslinking assay of VDAC1 2 h after lesion with 0.03%, 0.05% and 0.07% paraformaldehyde (PFA) concentrations, in which it was possible to identify monomers (i), dimers (ii), and other oligomers (iii and iv). K VDAC1 protein in mitochondria, plasma membrane, and endoplasmic reticulum in controls and 2 h after the lesion. Graph of normalized optical density (OD) of the subcellular fractionation experiment. Scale bar: 50 μm. All analyses considered n = 6 and *P < 0.05 in paired Student’s t-test. ONL outer nuclear layer, OPL outer plexiform layer, INL inner nuclear layer, IPL inner plexiform layer, GCL ganglion cell layer, The illustrations were created using BioRender software.

Fig 3: Overexpression of ERP44 attenuates H/R-induced cardiomyocyte pyroptosis.A CCK-8 was used to detect cell viability; B colorimetric assay was used to detect Caspase-1 activity; C western blot was used to detect the levels of pyroptosis proteins; D ELISA was used to detect the levels of IL-1β and IL-18. Nif represents calcium channel blocker Nifedipine D6. Compared to the H/R group, **p < 0.01, ***p < 0.001. All experiments were performed three times independently. Data in panel B were analyzed using one-way ANOVA and data in panels A/C/D were analyzed using two-way ANOVA. Tukey’s multiple comparison test was applied for the post hoc test after ANOVA.