Journal: Inflammation research : official journal of the European Histamine Research Society ... [et al.]

Article Title: A positive feedback cycle between the alarmin S100A8/A9 and NLRP3 inflammasome-GSDMD signalling reinforces the innate immune response in Candida albicans keratitis.

doi: 10.1007/s00011-023-01757-5

Figure Lengend Snippet: Fig. 6 Alarmin S100A8/A9 forms a positive feedback loop with NLRP3 inflammasome- GSDMD in the pathogenesis of Candida albicans keratitis

Article Snippet: After intraperitoneal injection system anaesthesia, mice were subconjunctivally injected with 10 μL of mS100A8 siRNA (50 nM, Ribobio, #siBDMV002), 10 μL of mNLRP3 siRNA (50 nM, Ribobio, #siBDMV002), 10 μL of mouse recombinant S100A8/A9 protein (1000 ng/mL, CloudClone, #RPK504Mu01), 10 μL of paquinimod (2 μg/μL, MCE, #ABR25757), or 10 μL of azeliragon (2 μg/μL, MCE, #HY50682) twice at 24 h and 6 h before C. albicans infection.

Techniques:

Journal: iScience

Article Title: Whitening of brown adipose tissue inhibits osteogenic differentiation via secretion of S100A8/A9

doi: 10.1016/j.isci.2024.108857

Figure Lengend Snippet: Loss of Rheb in BAT induces transcription and secretion of S100A8/A9 Interscapular BAT from either control or Rheb AD KO (A), or from control or Rheb BAD KO (B) mice, were used for transcriptome analysis by RNA-sequencing. The significantly upregulated genes were further subjected to gene ontology (GO) analysis. Detailed biological processes of the significantly upregulated genes in the Rheb AD KO mice are shown as a CNET plot (C). (D) Significantly up- or downregulated genes (at least two-fold, p < 0.01) in the BAT of control or Rheb BAD KO mice are shown as a volcano plot, with S100A8 and S100A9 labeled. S100A8 and S100A9 expression in the BAT of control or Rheb AD KO (or control or Rheb BAD KO) mice was analyzed by RT-qPCR (E) or western blotting (F). (G) Serum S100A8 and S100A9 levels of these mice were determined by ELISA or western blotting. Interscapular BAT (H) or serum (I) of the 3-month and 20-month C57BL/6J mice were analyzed for S100A8 expression by western blotting. (J) Conditional medium (CM) collected from interscapular BAT or bone marrow cells from these mice were analyzed for S100A8 by western blotting. Silver staining was used to assess equal loading. Data are shown as box-and-whisker plots (with median and interquartile ranges) from max to min, with all data points shown. Analyses were performed as two-tailed unpaired t-test.

Article Snippet: BMSC and C3H/10T1/2 were treated with 200, 500, and 1000 ng/mL of endotoxin-free S100A8/A9 RP (8916-S8-050, R&D, USA) for 7 days in osteogenic medium.

Techniques: RNA Sequencing Assay, Labeling, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Silver Staining, Whisker Assay, Two Tailed Test

Journal: iScience

Article Title: Whitening of brown adipose tissue inhibits osteogenic differentiation via secretion of S100A8/A9

doi: 10.1016/j.isci.2024.108857

Figure Lengend Snippet: Brown adipocytic S100A8/A9 mediates the effect of BAT Rheb loss on osteogenesis (A) Interscapular BAT from either control or Rheb AD KO, or from control or Rheb BAD KO mice, were analyzed for S100A8 or S100A9 expression by immunohistochemistry. The conditional medium (CM) was collected from both interscapular BADs and epididymal WADs of the control and Rheb AD KO mice (B), or from interscapular BADs of the control and Rheb BAD KO mice (C). The cells were then lysed, followed by both the whole-cell lysates and CM analyzed for S100A8 and S100A9 expression by WB. Primary brown adipocyte progenitors (BPADs) from wild-type C57BL/6J mice were transfected with lentivirus carrying the control or Rheb shRNA and induced into mature BADs, followed by analysis of whole-cell or secreted S100A8 level via RT-qPCR (D) or WB (E), respectively. BPADs were induced into BADs and treated with various concentrations of rapamycin for 12 h, then analyzed for S100A8 expression via RT-qPCR (F) and WB (G). The efficiency of rapamycin treatment was assessed by monitoring phosphorylated S6 (S235/236) (G). S100A8/A9 neutralizing antibody was injected into the tibial medullary cavity of Rheb BAD KO mice (10-month) every 3 days for 2 months, and the effects on bone formation were analyzed by 3D-microCT (H) or HE staining (I). Bone histomorphometric parameters of the tibiae were shown in the lower panel of (H). Data are shown as box-and-whisker plots (with median and interquartile ranges) from max to min, with all data points shown. Two-tailed unpaired t-test was used for two-group comparison; for comparison between multiple groups, one-way analysis of variance with multiple comparisons were used, followed by the Bonferroni post-hoc test for significance.

Article Snippet: BMSC and C3H/10T1/2 were treated with 200, 500, and 1000 ng/mL of endotoxin-free S100A8/A9 RP (8916-S8-050, R&D, USA) for 7 days in osteogenic medium.

Techniques: Expressing, Immunohistochemistry, Transfection, shRNA, Quantitative RT-PCR, Injection, Staining, Whisker Assay, Two Tailed Test, Comparison

Journal: iScience

Article Title: Whitening of brown adipose tissue inhibits osteogenic differentiation via secretion of S100A8/A9

doi: 10.1016/j.isci.2024.108857

Figure Lengend Snippet: S100A8/A9 inhibits OB differentiation of BMSCs through targeting toll-like receptor 4 (TLR4) (A) Expression of the S100A8/A9 receptor TLR4 and RAGE in primary BMSCs was analyzed by RT-qPCR. C2C12 stromal cells (B) and C3H10T1/2 MSCs (C) were treated with recombinant S100A8/A9 and analyzed for Myd88 expression and localization by confocal microscopy. Original magnification, ×200 or ×600; scale bar was shown as indicated. (D) C3H10T1/2 cells were treated with S100A8/A9 and TLR4 inhibitor TAK242 as indicated, followed by assay for OB differentiation by ALP staining (upper panel) and WB (lower panel). For comparisons between 2 groups, two-tailed unpaired t-tests were used. For comparisons between multiple groups, one-way analysis of variance with multiple comparisons were used, followed by the Bonferroni post-hoc test for significance. Data are shown as box-and-whisker plots (with median and interquartile ranges) from max to min, with all data points shown. Two-tailed unpaired t-test was used for two-group comparison; for comparison between multiple groups, one-way analysis of variance with multiple comparisons were used, followed by the Bonferroni post-hoc test for significance.

Article Snippet: BMSC and C3H/10T1/2 were treated with 200, 500, and 1000 ng/mL of endotoxin-free S100A8/A9 RP (8916-S8-050, R&D, USA) for 7 days in osteogenic medium.

Techniques: Expressing, Quantitative RT-PCR, Recombinant, Confocal Microscopy, Staining, Two Tailed Test, Whisker Assay, Comparison

Journal: iScience

Article Title: Whitening of brown adipose tissue inhibits osteogenic differentiation via secretion of S100A8/A9

doi: 10.1016/j.isci.2024.108857

Figure Lengend Snippet:

Article Snippet: BMSC and C3H/10T1/2 were treated with 200, 500, and 1000 ng/mL of endotoxin-free S100A8/A9 RP (8916-S8-050, R&D, USA) for 7 days in osteogenic medium.

Techniques: Virus, Negative Control, Recombinant, Staining, Silver Staining, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Real-time Polymerase Chain Reaction, Software