primary antibodies against cleaved-gsdmd  (Cell Signaling Technology Inc)

Journal: European Journal of Medical Research

Article Title: High-fat diet activates pyroptosis of retinal pigment epithelial cells in aged TgAPPswePS1 transgenic mice

doi: 10.1186/s40001-025-02898-5

Figure Lengend Snippet: Immunofluorescence analysis of the pyroptosis-related proteins in RPE cells. Scale bar = 20 μm. A Representative images showing the expressions of NLRP3, caspase-1 and GSDMD in RPE cells. B Representative images showing the expressions of IL-1β and IL-18 in RPE cells. C Semi-quantitative analysis of fluorescence intensity for NLRP3, caspase-1, GSDMD, IL-1β and IL-18 expressions in RPE cells. ** p < 0.01, ***p < 0.001, ****p < 0.0001, ns, no significance

Article Snippet: After blocking with 5% nonfat dry milk, membranes were incubated overnight at 4 °C with primary antibodies against NLRP3 (1:1000, #15101), Pro-Caspase-1 (1:1000, #24232), Cleaved-Caspase-1 (1:1000, #89332), Cleaved-GSDMD (1:1000, #10137, Cell Signaling Technology, MA, USA), GSDMD (1:1000, ab219800) and β-actin (1:1000, ab8226, Abcam Biotechnology, UK).

Techniques: Immunofluorescence, Fluorescence

Journal: European Journal of Medical Research

Article Title: High-fat diet activates pyroptosis of retinal pigment epithelial cells in aged TgAPPswePS1 transgenic mice

doi: 10.1186/s40001-025-02898-5

Figure Lengend Snippet: Expression alternations of pyroptosis-related proteins in RPE cells. A NLRP3, caspase-1, GSDMD protein expressions in RPE were detected via western blotting. B IL-1β and IL-18 levels in RPE were tested via ELISA. * p < 0.05, **p < 0.01, ***p < 0.001, ns, no significance

Article Snippet: After blocking with 5% nonfat dry milk, membranes were incubated overnight at 4 °C with primary antibodies against NLRP3 (1:1000, #15101), Pro-Caspase-1 (1:1000, #24232), Cleaved-Caspase-1 (1:1000, #89332), Cleaved-GSDMD (1:1000, #10137, Cell Signaling Technology, MA, USA), GSDMD (1:1000, ab219800) and β-actin (1:1000, ab8226, Abcam Biotechnology, UK).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Annals of Translational Medicine

Article Title: Interleukin-35 attenuates blood-brain barrier dysfunction caused by cerebral ischemia-reperfusion injury through inhibiting brain endothelial cell injury

doi: 10.21037/atm-22-2770

Figure Lengend Snippet: IL-35 suppressed OGD/R-induced ROS generation in endothelial cells. The effects of IL-35 on ROS generation and the MAPK pathway in bEnd.3 cells after 6 hours of OGD and 18 hours reoxygenation. (A) Intracellular ROS was viewed with fluorescence microscopy (scale bar: 400 µm). (B) The resulting fluorescence intensity was quantified using Image J software. Error bars represent mean ± SD of at least three independent experiments. (C) The concentration of MDA in cell lysates. (D-F) Representative Western blot bands and quantitative analysis of the ratio of p-p38/p38 and TXNIP. The band intensities were measured using scanning densitometry. Protein expressions were normalized to β-actin (n=3). Data are expressed as mean ± SD. PBS, phosphate buffered saline; OGD-R, oxygen-glucose deprivation and reperfusion; IL, interleukin; IOD, integrated option density; MDA, malondialdehyde; TXNIP, thioredoxin-interacting protein; ROS, reactive oxygen species; MAPK, mitogen-activated proliferation protein kinase; SD, standard deviation.

Article Snippet: After blocking with 5% nonfat dry milk, the membranes were incubated overnight with primary antibodies against ZO-1, occludin, cleaved caspase-1, GSDMD, and TXNIP (1:1,000, Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (1:5,000, Yi Fei Xue biotechnology, Nanjing, China).

Techniques: Fluorescence, Microscopy, Software, Concentration Assay, Western Blot, Saline, Standard Deviation

Journal: Annals of Translational Medicine

Article Title: Interleukin-35 attenuates blood-brain barrier dysfunction caused by cerebral ischemia-reperfusion injury through inhibiting brain endothelial cell injury

doi: 10.21037/atm-22-2770

Figure Lengend Snippet: IL-35 protected OGD/R-induced endothelial cell injury. bEnd.3 cells were pretreated with or without IL-35 and then subjected to 6 hours of OGD and 18 hours reoxygenation. (A-C) Western blot images and analysis of cleaved caspase-1 and GSDMD expressions in bEnd.3 cells. (D,E) Results of ELISA showing the levels of IL-18 and IL-1β in the culture supernatant. (F-H) Results of Western blot assay showing the protein expressions of ZO-1 and occludin in bEnd.3 cells. Error bars represent mean ± SD of at least three independent experiments. GSDMD, gasdermin D; PBS, phosphate buffered saline; OGD-R, oxygen-glucose deprivation and reperfusion; IL, interleukin; ZO-1, zonula occludens-1; bEnd.3, brain endothelial cells; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation.

Article Snippet: After blocking with 5% nonfat dry milk, the membranes were incubated overnight with primary antibodies against ZO-1, occludin, cleaved caspase-1, GSDMD, and TXNIP (1:1,000, Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (1:5,000, Yi Fei Xue biotechnology, Nanjing, China).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation