Journal: Shock (Augusta, Ga.)

Article Title: TARGETING S100A9-TLR2 AXIS CONTROLS MACROPHAGE NLRP3 INFLAMMASOME ACTIVATION IN FATTY LIVER ISCHEMIA REPERFUSION INJURY

doi: 10.1097/SHK.0000000000002470

Figure Lengend Snippet: IR-induced injury leads to increased S100A9 and related pathway activation in fatty livers HFD fed C57 mice were used to establish liver IR models. Samples were harvested after 90-min ischemia and 6 h of reperfusion. (A) Serum ALT levels. (B) Immunofluorescence staining and quantification of CD11b + macrophages in ischemia fatty livers, scale bars: 50 μm, 20 μm. (C) Volcano Plot displaying 748 genes to be upregulated and 625 genes downregulated. (D) KEGG pathway enrichment analysis of major biological pathways contributing to fatty liver IR stress. (E) Heatmap showing the expression of different genes. (F) Western-assisted analysis and relative density ratio of S100A9, TLR2, and ATF4 in liver tissues. (G) Immunochemistry staining and quantification of S100A9 positive cells in ischemia fatty livers, scale bars: 50 μm, 20 μm. (H) Detection of cytokines IL-1β, CXCL1, CXCL-2, and IL-6 by qRT-PCR in the livers. N = 3–6/group. All data represent the mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001. ALT, alanine aminotransferase; HFD, high-fat diet; KEGG, Kyoto Encyclopedia of Genes and Genomes; IR, ischemia reperfusion.

Article Snippet: For immunofluorescence (IF) staining, liver macrophages were detected using primary CD11b mouse mAb (65055-1-Ig, Proteintech, Wuhan, China).

Techniques: Activation Assay, Immunofluorescence, Staining, Expressing, Western Blot, Quantitative RT-PCR

Journal: Shock (Augusta, Ga.)

Article Title: TARGETING S100A9-TLR2 AXIS CONTROLS MACROPHAGE NLRP3 INFLAMMASOME ACTIVATION IN FATTY LIVER ISCHEMIA REPERFUSION INJURY

doi: 10.1097/SHK.0000000000002470

Figure Lengend Snippet: S100A9 knockdown attenuates liver dysfunction and inflammatory damage following IR of fatty livers C57 mice were injected with AAV9-S100A9-shRNA 48 h before establishing liver IR models using HFD-fed mice. Samples were harvested after 90-min ischemia and 6 h of reperfusion. (A) Western-assisted analysis and relative density ratio of S100A9 in liver tissues. (B) H&E staining of ischemic fatty livers, scale bar: 200 μm, 100 μm. (C) Serum ALT levels. (D) Immunofluorescence staining and quantification of CD11b positive macrophages and immunochemistry staining and quantification of Ly6G positive cells in ischemia fatty livers, scale bar: 40 μm. (E) Western-assisted analysis of TLR2 and ATF4 in liver tissues. (F) Dual immunofluoresence staining for CD68 (red) and TLR2 (green) co-localization in injured fatty livers after IR stimulation, scale bars: 50 μm, 20 μm. (G) Detection of cytokines IL-1β, CXCL1, CXCL-2, and IL-6 by qRT-PCR in the livers. N = 3–6/group. All data represent the mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001. HFD, high-fat diet; IR, ischemia reperfusion. qRT-PCR, quantitative real-time PCR.

Article Snippet: For immunofluorescence (IF) staining, liver macrophages were detected using primary CD11b mouse mAb (65055-1-Ig, Proteintech, Wuhan, China).

Techniques: Knockdown, Injection, shRNA, Western Blot, Staining, Immunofluorescence, Quantitative RT-PCR, Real-time Polymerase Chain Reaction