q zhang  (ATCC)

Journal: Cellular signalling

Article Title: The cGAS/STING signaling pathway is involved in sevoflurane induced neuronal necroptosis via regulating microglia M1 polarization.

doi: 10.1016/j.cellsig.2024.111195

Figure Lengend Snippet: Fig. 2. Identification of cGAS/STING from RNA sequencing analysis of the sevoflurane and control groups. Volcano plot of differentially expressed genes. B. Heatmap of significantly differentially expressed genes. C. Gene Ontology (GO) analysis of differentially expressed genes. D. Kyoto Encyclopedia of Genes and Genomes analysis of differentially expressed genes. E. GO analysis of 20 upregulated differentially expressed genes. F. Western blotting was performed to detect cGAS,/STING expression under sevoflurane exposure. G–H. Quantitative analysis of cGAS and STING. Data are expressed as the mean ± SD, *P < 0.05, **P < 0.01.

Article Snippet: After blocking with 5% bovine serum albumin in Tris-Cl buffer containing 0.1% Tween-20 for 1 h at room temperature, the membrane was incubated with primary antibodies against inducible nitric oxide synthase (iNOS, Cat. 18,985–1-AP, Proteintech), CD16 (Cat. A23540, ABclonal), CD32 (Cat. 15,625–1-AP, Proteintech), receptorinteracting protein kinase 3 (RIPK3, Cat. 17,563–1-AP, Proteintech), phosphorylated mixed lineage kinase domain-like (p-MLKL, Cat. ET1705–51, HUABIO), RIPK1 (Cat. 17,519–1-AP, Proteintech), cGAS Q. Zhang et al. Cellular Signalling 119 (2024) 111195 (Cat. PA5–56820, Proteintech), phosphorylated STING (p-STING, Cat. PA5–105674, Proteintech), STING (Cat. PA5–23381, Proteintech), NFκB (Cat. ab207297, Abcam), IκB (Cat. #4814, CellSignaling Technology), and β-actin (Cat. AC021, ABclonal) overnight at 4 ◦C.

Techniques: RNA Sequencing, Control, Western Blot, Expressing

Journal: Cellular signalling

Article Title: The cGAS/STING signaling pathway is involved in sevoflurane induced neuronal necroptosis via regulating microglia M1 polarization.

doi: 10.1016/j.cellsig.2024.111195

Figure Lengend Snippet: Fig. 3. STING knockdown inhibits amoebic morphology and pro-inflammatory M1 polarization in microglia. A. Western blotting was used to detect STING after exposure to three different STING siRNAs: STING siRNA-865, STING siRNA-1036, and STING siRNA-1262. B. Quantitative analysis of STING. C. Photograph of BV2 microglial cells with STING knockdown under sevoflurane exposure. D. Western blotting was used to detect inducible nitric oxide synthase (iNOS), cGAS, STING, CD16, and CD32. E–I. Quantitative analysis of iNOS, cGAS, STING, CD16, and CD32, separately. J. An enzyme- linked immunosorbent assay was used to detect tumor necrosis factor-α in the cell supernatant. Data are expressed as the mean ± SD, *P < 0.05, **P < 0.01.

Article Snippet: After blocking with 5% bovine serum albumin in Tris-Cl buffer containing 0.1% Tween-20 for 1 h at room temperature, the membrane was incubated with primary antibodies against inducible nitric oxide synthase (iNOS, Cat. 18,985–1-AP, Proteintech), CD16 (Cat. A23540, ABclonal), CD32 (Cat. 15,625–1-AP, Proteintech), receptorinteracting protein kinase 3 (RIPK3, Cat. 17,563–1-AP, Proteintech), phosphorylated mixed lineage kinase domain-like (p-MLKL, Cat. ET1705–51, HUABIO), RIPK1 (Cat. 17,519–1-AP, Proteintech), cGAS Q. Zhang et al. Cellular Signalling 119 (2024) 111195 (Cat. PA5–56820, Proteintech), phosphorylated STING (p-STING, Cat. PA5–105674, Proteintech), STING (Cat. PA5–23381, Proteintech), NFκB (Cat. ab207297, Abcam), IκB (Cat. #4814, CellSignaling Technology), and β-actin (Cat. AC021, ABclonal) overnight at 4 ◦C.

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

Journal: Cellular signalling

Article Title: The cGAS/STING signaling pathway is involved in sevoflurane induced neuronal necroptosis via regulating microglia M1 polarization.

doi: 10.1016/j.cellsig.2024.111195

Figure Lengend Snippet: Fig. 5. Schematic of the role of cGAS/STING signaling pathway in sevoflurane induced neuronal necroptosis via regulating microglia M1 polarization. Sevoflurane facilitated microglial M1 polarization through the cGAS/STING signaling pathway, resulting in increased release of inflammatory factors such as tumor necrosis factor-α and hastening neuronal necroptosis triggered by calcium overload.

Article Snippet: After blocking with 5% bovine serum albumin in Tris-Cl buffer containing 0.1% Tween-20 for 1 h at room temperature, the membrane was incubated with primary antibodies against inducible nitric oxide synthase (iNOS, Cat. 18,985–1-AP, Proteintech), CD16 (Cat. A23540, ABclonal), CD32 (Cat. 15,625–1-AP, Proteintech), receptorinteracting protein kinase 3 (RIPK3, Cat. 17,563–1-AP, Proteintech), phosphorylated mixed lineage kinase domain-like (p-MLKL, Cat. ET1705–51, HUABIO), RIPK1 (Cat. 17,519–1-AP, Proteintech), cGAS Q. Zhang et al. Cellular Signalling 119 (2024) 111195 (Cat. PA5–56820, Proteintech), phosphorylated STING (p-STING, Cat. PA5–105674, Proteintech), STING (Cat. PA5–23381, Proteintech), NFκB (Cat. ab207297, Abcam), IκB (Cat. #4814, CellSignaling Technology), and β-actin (Cat. AC021, ABclonal) overnight at 4 ◦C.

Techniques: