Fig. 6: GRP75 silencing ameliorates ZBP1 activation and PANoptosis. | Communications Biology

Fig. 6: GRP75 silencing ameliorates ZBP1 activation and PANoptosis.

From: Endoplasmic reticulum-mitochondria coupling prompts ZBP1-mediated RPE cell PANoptosis in age-related macular degeneration

Fig. 6: GRP75 silencing ameliorates ZBP1 activation and PANoptosis.The alternative text for this image may have been generated using AI.

ARPE-19 cells were transfected with GRP75 siRNA under NaIO3 (10 mM, 24 h) stimulation. A Immunofluorescence staining reveals that mPTP opening in ARPE-19 cells were decreased significantly by GRP75 siRNA. Scale bars = 25 μm. B qRT-PCR assay showing that elevated cytoplasmic mtDNA (MTND1, MTND2, ATP6, CO2, DLOOP) level stimulated by NaIO3 were reversed after treatment with GRP75 siRNA (n = 3). Immunofluorescence staining showing that NaIO3 induces C relocation of dsDNA (anti-dsDNA, green) from mitochondria (anti-TOMM20, red) into cytoplasm and D sensed by ZBP1 (anti-ZBP1, red). Nuclei were stained with DAPI. GRP75 siRNA decreased C the dsDNA leakage and D colocalization of dsDNA with ZBP1. Scale bars = 25 μm. E Calcein-AM/PI assay reveals dead (red) and live (green) ARPE-19 cells. Scale bars = 50 μm. F CCK-8 assay showing ARPE-19 cells viability (n = 6). G PANoptosis-related protein expressions (p-MLKL, MLKL, cleaved-GSDMD, GSDMD, cleaved-Caspase3, Caspase3) in primary mouse RPE cells by immunoblots analysis. H RPE65 and ZO-1 expressions in primary mouse RPE cells by immunoblots analysis. I ARPE-19 cells stained with ZO-1 (green) and DAPI (blue) showing ARPE-19 cells dysfunction treated with NaIO3 and was reversed significantly by GRP75 siRNA. Scale bars = 25 μm. Data are mean ± SD, the p values were obtained using one-way ANNOVA (F–H) or unpaired t-test (B). *p < 0.05 versus control group, #p < 0.05 versus NC group.

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