Fig 1: Inflammasome activation led to pyroptosis in ZIKV-infected macrophages.Cell lysates, prepared at various time points p.i. from THP-1 (A) and RAW264.7 (B) cells infected with 0.1 MOI of ZIKV, were analysed by western blot analyses with antibodies for pro- or cleaved caspase-1 and GSDMD. Quantitative analyses of the grayscale values in the blots of the pro- and cleaved caspase-1 and GSDMD in infected and control THP-1 (C, D, & F) and RAW264.7 (E, G, & H) cells were shown. The experiments were performed in triplicates and the data were shown as mean+SD, analyzed by unpaired Students t-test. *, P<0.05; **, P<0.01.
Fig 2: TLR4/ NF-?B and GSDMD-NT expressions were increased in HK-2 cells under HG condition. (A) WB and densitometric analysis of TLR4 in HK-2 cells exposed to different concentrations of D-glucose. (B) Protein expression of GSDMD-NT exposed to different concentrations of D-glucose. The protein level of TLR4 (A1) and GSDMD-NT(B1) in HK-2 cells exposured to D-glucose (5.5, 17.5, 30, and 45 mM) for 24 h. Each assay was representative of three independent experiments. Data were expressed as means ±SEM. (*P < 0.05 vs. control; **P < 0.05 vs. 17.5Glu group; ***P < 0.05 vs. 30Glu group).
Fig 3: TLR4-induced pyroptosis in vivo relies on lncRNA-F630028O10Rik.a LncRNA-F630028O10Rik levels in TLR4-/- mice injected with plvxF60 after SCI. b NLRP3, ASC, CASP-1 and GSDMD mRNA levels in TLR4-/- mice with/out plvxF60 treatment. c, d IL-1b, IL-6, TNF-a and LDH levels in the spinal cord homogenates of the above mice. e BBB scores of the different groups at the designated time points. f The negative control of IHC. Rabbit serum and PBS were used to instead of primary antibodies to exclude non-specific staining. g Representative HE and IHC images of NLRP3 and GSDMD in each group. The immunohistochemical staining was conducted in spinal cord sections at the epicenter and peri-lesion sites. Scale bar: 50 µm. *p < 0.05, **p < 0.01, ***p < 0.001 versus WT, #p < 0.05, ##p < 0.01, ###p < 0.001 versus TLR4-/-+plvxF60.Data are the mean ± SD from 3 independent experiments.
Fig 4: Western blot bands of the expression of NLRP3, ASC, GSDMD, and caspase-1/3/4/5 after the different treatments. Data are expressed as mean ± SEM (n = 3). *P > 0.05 compared with blank control; #P < 0.05 compared with the rabbit group. Blank control, blank serum control group; rabbit control, rabbit serum control group; JWJGC-H, JWJGC high-dose group; JWJGC-M, JWJGC medium-dose group; JWJGC-L, JWJGC low-dose group; Leflunomide, leflunomide-positive control group.
Fig 5: HBV X protein (HBx) promoted the upregulation and activation of NLRP3 inflammasome expression under H2O2 stress. Quantification of mRNA expression of GAPDH, NLRP3, ASC, caspase-1, IL-1ß, IL-18, HMGB1, and GSDMD using real-time PCR, n = 5 (a, b). Data are expressed as mean ± SD. *P < 0.05 and **P < 0.01. Western blot analysis of NLRP3, ASC, pro-caspase-1, Cleaved-caspase-1 (p10), and IL-1ß (mature IL-1ß), n = 4 (c, d). GAPDH served as a loading control
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