DFNA5/GSDME Antibody - N-terminal - #AF4016

Fig. 6 (A-D) Intracellular ROS levels were measured by CLSM stained with DCFH-DA and analyzed by FCM. Scale bars: 100 μm. (E) ROS images and pyroptotic morphological changes were captured by CLSM. Vitamin C (Vc) and vitamin K3 (Vk3) were used as a 1O2 scavenger and 1O2 inducer, respectively; scale bar: 40 μm. The yellow arrows represent the pyroptotic bodies. (F) Intracellular ROS levels were measured by FCS after pretreatment with Vc or Vk3. (G) Cleavage of GSDME and caspase-3 from CT26 cells following 8-hour treatment of CUR@PLGA-NPs + Light at a concentration of 10 µM as detected by western blot analysis. (H) CLSM images showing GSDME-N expression after being treated with Control, CuET, Cisplatin, NPs + Light, CuET + NPs + Light, and Cisplatin + NPs + Light. (I) LDH release analysis showing influence of CuET and Cisplatin on NPs + Light-induced pyroptosis. (J) Schematic illustration of the proposed pyroptosis activation via 1O2 generation

Fig. 6 (A-D) Intracellular ROS levels were measured by CLSM stained with DCFH-DA and analyzed by FCM. Scale bars: 100 μm. (E) ROS images and pyroptotic morphological changes were captured by CLSM. Vitamin C (Vc) and vitamin K3 (Vk3) were used as a 1O2 scavenger and 1O2 inducer, respectively; scale bar: 40 μm. The yellow arrows represent the pyroptotic bodies. (F) Intracellular ROS levels were measured by FCS after pretreatment with Vc or Vk3. (G) Cleavage of GSDME and caspase-3 from CT26 cells following 8-hour treatment of CUR@PLGA-NPs + Light at a concentration of 10 µM as detected by western blot analysis. (H) CLSM images showing GSDME-N expression after being treated with Control, CuET, Cisplatin, NPs + Light, CuET + NPs + Light, and Cisplatin + NPs + Light. (I) LDH release analysis showing influence of CuET and Cisplatin on NPs + Light-induced pyroptosis. (J) Schematic illustration of the proposed pyroptosis activation via 1O2 generation

Fig. 3 Pyroptosis induced by iron accumulation may occur in splenic B cells after SCI. a Three days after SCI, serum samples were collected for quantification of the protein expression levels of MCP-1, IL-1β, IL-6, and TNF-α by ELISA. b Three days after SCI, injured spinal cord homogenates were collected for quantification of the protein expression levels of MCP-1, IL-1β, IL-6, and TNF-α by ELISA. c Three days after SCI, the spleen was stained with Prussian blue to detect the level of iron ion. d Three days after SCI, serum samples were collected to quantify the concentration of iron ions. e Three days after SCI, B cell lysates were collected to quantify the concentration of iron ions. f Detection of Tom20-Bax-caspase-GSDME pathway-related protein expression in B cells by western blot. g The knock-down efficiency of AAV on Tom20 in B cells was verified. h, l, m The expression levels of MCP-1, IL-1 β, IL-6, TNF- α, IgG and IgM in serum of different groups were detected by ELISA. i–k Three days after SCI, the spleens of mice were observed, and the spleen length and organ index of different groups were compared. n, o, q The spleen was taken for HE staining, Prussian blue staining, and immunofluorescence. CD19+ was red, dapi was blue, and merge was combined. p Detection of nerve evoked potentials in different groups of mice. r Detection of the expression of Tom20-Bax-caspsae-GSDME pathway-related proteins in different groups of B cells. All data are expressed as the mean ± SD (n ≥ 3 replicates per group). ns P > 0.05, * P 

Fig. 2. MPPa-PDT induced pyroptosis in human osteosarcoma 143b and HOS cells. (A) Morphological changes of the cells, the red arrow indicated pyroptosis cells. (B) The ultrastructure of cells was observed with an SEM. (C) GSDME-FL and GSDME-NT levels were analyzed using Western blotting. *: p < 0.05 vs. Control group.

Fig. 1. Expression of GSDME and GSDME-N after 6–72 h of reperfusion in mouse MCAO model. (A) The expression of GSDME mRNA in cerebral ischemic penumbra tissue was detected by RT-PCR; (B) GSDME and GSDME-N protein expression in cerebral ischemic penumbra tissue was analyzed by Western blot. Data were shown as the means ± SEM, n = 5 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.