Journal: Oxidative Medicine and Cellular Longevity

Article Title: Recurrent Hypoglycemia Impaired Vascular Function in Advanced T2DM Rats by Inducing Pyroptosis

doi: 10.1155/2022/7812407

Figure Lengend Snippet: NF- κ B, NLRP3, and cGAS–STING pathway activity of the aortas in response to acute and recurrent hypoglycemia in aged T2DM rats. (a) p-p65, NLRP3, ASC, (b) cleaved caspase-1, cGAS, and (e) STING expression were assayed by western blotting. The expression and location of NLRP3 were determined by (c) immunohistochemistry and (d) immunofluorescence; ∗ p < 0.05 DM vs. control; # p < 0.05 H-DM, RH-DM vs. DM; & p < 0.05 H-DM vs. RH-DM.

Article Snippet: Proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis and incubated with primary antibodies: eNOS (1 : 1000, ab300071, Abcam), iNOS (1 : 1000, ab283655, Abcam), NOX2 (1 : 2000, 19013-1-AP, ProteinTech), NOX4 (1 : 2000, 14347-1-AP, ProteinTech), p-p65 (1 : 1000, 3033, Cell Signaling Technology), p65 (1 : 1000, 8242, Cell Signaling Technology), NLRP3 (1 : 1000, NBP2-12446, NOVUS), ASC (1 : 1000, sc-514414, Santa Cruz), Caspase-1 (1 : 1000, bs-10743R, Bioss), cGAS (1 : 1000, NBP3-16666, NOVUS), STING (1 : 1000, CST50494, Cell Signaling Technology), GSDMD (1 : 1000, NBP2-33422, NOVUS), Bax (1 : 1000, 50599-2-Ig, ProteinTech), Bcl-2 (1 : 1000, 26593-1-AP, ProteinTech), and β -actin (1 : 5000, 20536-1-AP, ProteinTech).

Techniques: Activity Assay, Expressing, Western Blot, Immunohistochemistry, Immunofluorescence

Journal: Oxidative medicine and cellular longevity

Article Title: Hemoglobin Derived from Subarachnoid Hemorrhage-Induced Pyroptosis of Neural Stem Cells via ROS/NLRP3/GSDMD Pathway.

doi: 10.1155/2023/4383332

Figure Lengend Snippet: Figure 2: NLRP3 mediated pyroptosis in rat hippocampus after SAH. (a) Representative images of the rat brains in control and SAH group after operation. (b) The SAH grade scores of the rats in control and SAH groups. (c) IL-1β expression in hippocampus were detected by ELISA. (d) Representative IF images of NLRP3 and Caspase1 p20 in hippocampus of rats after SAH. Scale bar = 50 μm. Data were expressed as Mean ± SD (n = 6), ∗∗∗p < 0:001 vs. control group.

Article Snippet: Briefly, brain sections were fixed in 4% paraformaldehyde for approximately 24 h. Coronal paraffin-embedded 4μm thick slices were processed for antigen retrieval and blocked with 5% BSA for 1 h. After blocking, the slices were incubated overnight at 4°C with the following primary antibodies: mouse anti-nestin (1 : 300, Cell Signaling Technology, #4760); mouse anti-SOX2 (1 : 300, Abcam, #ab79351); rabbit anti-NLRP3 (1 : 400, Biorbyt, #orb1105906); rabbit anti-caspase1 p20 (1 : 400, Biorbyt, #orb454595); and rabbit anti-GSDMD (1 : 400, Biorbyt, #orb1105840).

Techniques: Control, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Oxidative medicine and cellular longevity

Article Title: Hemoglobin Derived from Subarachnoid Hemorrhage-Induced Pyroptosis of Neural Stem Cells via ROS/NLRP3/GSDMD Pathway.

doi: 10.1155/2023/4383332

Figure Lengend Snippet: Figure 4: The NLRP3/GSDMD mediated the pyroptosis in primary NSCs exposed to Hb. (a) Hb treatment induced the elevated mRNA expressions of NLRP3, ASC, and Caspase1 in NSCs using qRT-PCR assay. (b) Western blot analysis showed the upregulated protein expressions of NLRP3, ASC, Cleaved Caspase1, and Cleaved GSDMD in Hb-treated NSCs. The band densities in western blot analysis were quantified using Image J Software. (c) Representative IFs staining images of NLRP3,Caspase1 P20, and GSDMD in NSCs with or without Hb treatment at 24 h. Scale bars = 50 μm. Data were expressed as Mean ± SD (n = 4), ∗p < 0:05, ∗∗p < 0:01, ∗∗∗p < 0:001 vs. control group.

Article Snippet: Briefly, brain sections were fixed in 4% paraformaldehyde for approximately 24 h. Coronal paraffin-embedded 4μm thick slices were processed for antigen retrieval and blocked with 5% BSA for 1 h. After blocking, the slices were incubated overnight at 4°C with the following primary antibodies: mouse anti-nestin (1 : 300, Cell Signaling Technology, #4760); mouse anti-SOX2 (1 : 300, Abcam, #ab79351); rabbit anti-NLRP3 (1 : 400, Biorbyt, #orb1105906); rabbit anti-caspase1 p20 (1 : 400, Biorbyt, #orb454595); and rabbit anti-GSDMD (1 : 400, Biorbyt, #orb1105840).

Techniques: Quantitative RT-PCR, Western Blot, Software, Staining, Control