Journal: JCI Insight

Article Title: Caspase-4/11 promotes hyperlipidemia and chronic kidney disease–accelerated vascular inflammation by enhancing trained immunity

doi: 10.1172/jci.insight.177229

Figure Lengend Snippet: ( A and B ) Flow cytometry analysis demonstrated that HFD+CKD increased the recruitment of CD45 + CD11b + monocytes and CD45 + CD11b + F4/80 + macrophages into the WT aorta, which were significantly suppressed in HFD+CKD casp11 –/– aortas. ( C and D ) Flow cytometry analysis demonstrated that casp11 –/– decreased blood CD45 + CD11b + monocytes in HFD-sham mice compared with WT HFD-sham mice; and HFD+CKD mice did not significantly change blood CD45 + CD11b + monocytes and CD45 + CD11b + F4/80 + macrophages in HFD+CKD mice ( n = 6–8). Flow cytometry analysis showed the infiltrated monocytes (CD11b + CD45 + ) and macrophages (CD11b + CD45 + F4/80 + ) in the blood of WT and casp11 –/– HFD+CKD and HFD-Sham mice. ( E ) Cytokine array showed that casp11 –/– decreased HFD+CKD-induced chemokines and cytokines, including CCL2 and CCL22, MMP-3, chinitianase 3-like 1 (CHIL3L1), IL-12p40, myeloperoxidase, TNFRSF11b, and PCSK9 in plasma. Each sample was pooled from 3 mice in each group ( n = 3). ImageJ was used to quantify the bands, and the significantly changed proteins were indicated. The Kruskal-Wallis test with Benjamini and Hochberg multiple-comparison method was used to control the overall FDR of 5% ( A – E ).

Article Snippet: Flow cytometry antibodies were bought from BD Biosciences except for anti–GSDMD-PE (Santa Cruz, H11), anti–VCAM-1 PECy7 (BioLegend, 105720), anti-CD11b BUV395 (BD Biosciences, 563553), anti-CD45 APC Cy7 (BioLegend, catalog 103116), anti-CD31 BUV737 (BD Biosciences, 612802), and anti-F4/80 Alexa Fluor 647 (BD Pharmingen, catalog 565853).

Techniques: Flow Cytometry, Comparison, Control

Journal: iScience

Article Title: Protective effects of S100A8 on sepsis mortality: Links to sepsis risk in obesity and diabetes

doi: 10.1016/j.isci.2022.105662

Figure Lengend Snippet: Myelomonocytic cell-specific S100A8 knockout mice were vulnerable to lethal endotoxemia, and exogenous S100A8 supplementation it was protective Myelomonocytic cell-specific S100A8 knockout (Lyz2 cre :S100A8 floxed/floxed ) mice or S100A8 floxed/floxed mice at 10 weeks of age were used for experiments. (A) The mRNA expression of S100a8 in peritoneal macrophages and the lungs, spleen, pancreas, fat, inguinal lymph nodes, kidneys, and liver (n = 3). (B) The percentage of Ly6G or F4/80 positive cells in peritoneal cells and the mean fluorescence intensity (MFI) of PE (Ly6G) and FITC (F4/80) staining were analyzed by flow cytometry with anti-Ly6G-PE and anti-F4/80-Alexa488 antibodies (n = 6). (C) mRNA expression of the indicated inflammatory cytokine and S100 genes in peritoneal macrophages incubated with PBS or LPS (2 μg/mL) for 2 h (n = 3). (D) mRNA expression of the indicated inflammatory cytokine and S100 genes in peritoneal cells at 4 h after LPS (2 μg/mL) injection (n = 3). (E) Serum S100A8 levels at 4 h after PBS or LPS (12.5 μg/gBW) injection (n = 3). (F) The survival rates, (G) body temperature, and (H) blood glucose levels of mice injected with either PBS or S100A8 (0.1 μg/gBW) at 1 h after LPS (12.5 μg/gBW) stimulation were measured at 12, 24, 36, and 48 h (n = 5–8). All data from three or four independent experiments are represented as the mean ± SEM ∗p < 0.05, ∗∗p < 0.01; ## p < 0.01 vs. Lyz2 cre :S100A8 floxed/floxed (LPS + S100A8).

Article Snippet: Peritoneal cells from myelomonocytic cell-specific S100A8 KO mice and control mice were stained with an Alexa Fluor 488-labeled anti-F4/80 (6F12) antibody (BD Biosciences) or a PE-labeled anti-Ly6G (RB6-8C5) antibody (BD Biosciences).

Techniques: Knock-Out, Expressing, Fluorescence, Staining, Flow Cytometry, Incubation, Injection

Journal: iScience

Article Title: Protective effects of S100A8 on sepsis mortality: Links to sepsis risk in obesity and diabetes

doi: 10.1016/j.isci.2022.105662

Figure Lengend Snippet:

Article Snippet: Peritoneal cells from myelomonocytic cell-specific S100A8 KO mice and control mice were stained with an Alexa Fluor 488-labeled anti-F4/80 (6F12) antibody (BD Biosciences) or a PE-labeled anti-Ly6G (RB6-8C5) antibody (BD Biosciences).

Techniques: Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Cholesterol Assay, Software, Real-time Polymerase Chain Reaction