Journal: PLOS Pathogens

Article Title: Fungal chitin-binding glycoprotein induces Dectin-2-mediated allergic airway inflammation synergistically with chitin

doi: 10.1371/journal.ppat.1011878

Figure Lengend Snippet: (A) Bone marrow-derived dendritic cells (BMDCs) from C57BL/6 mice were incubated with recombinant LdpA (rLdpA) (1 μg/mL), chitin (50 μg/mL), rLdpA–chitin complex (rLdpA, 1 μg/mL; chitin, 50 μg/mL), lipopolysaccharide (LPS) (100 ng/mL), and phosphate-buffered saline (PBS) for 24 h. MHC class II, CD80, CD86, and CD40 cell-surface expression were measured by flow cytometry. (B) Proinflammatory cytokine and chemokine mRNA expression. After BMDCs derived from C57BL/6 mice were incubated with rLdpA (25 μg/mL), chitin (250 μg/mL), rLdpA–chitin complex (rLdpA, 1 μg/mL; chitin, 50 μg/mL), and PBS for 1.5 h, mRNA expression levels of Tnf-α , Il-1α , Il-1β , Il-12 p35 , Il-12 p40 , Il-6 , Kc/Cxcl1 , and Mip-2/Cxcl2 were measured by quantitative real-time PCR. mRNA expression levels were normalized relative to Gapdh mRNA levels and are shown as fold change relative to the control PBS group. (C, D) BMDCs derived from C57BL/6 mice were incubated with rLdpA (1 μg/mL), chitin (50 μg/mL), rLdpA–chitin complex (rLdpA, 1 μg/mL; chitin, 50 μg/mL), and PBS for 24 h, TNF-α, IL-1α, and KC/CXCL1 protein levels in the culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA) (C). Cytotoxicity was assessed by measuring the release of lactate dehydrogenase (LDH) (D). (E, F, G) BMDCs derived from C57BL/6, Card9 −/− , Myd88 −/− (E), Mincle −/− , MCL −/− (F), Dectin-2 −/− and Dectin-1 −/− (G) mice were incubated with rLdpA–chitin complex (rLdpA, 1 μg/mL; chitin, 50 μg/mL) and PBS for 24 h, and TNF-α, IL-1α, and KC/CXCL1 protein levels in the culture supernatant were measured by ELISA. Data are shown as the mean ± standard deviation (SD). * P < 0.05, ** P < 0.01, and *** P < 0.001 by one-way ANOVA with post hoc Tukey–Kramer test (B, C, D).

Article Snippet: Cells were stained with a mixture of fluorochrome-conjugated monoclonal antibodies (mAbs) to CD11c (N418; eBioscience), MHC class II (M5/114.15.2; Miltenyi Biotec), CD40 (FGK45.5; Miltenyi Biotec), CD80 (16-10A1; Miltenyi Biotec), and CD86 (PO3.3; Miltenyi Biotec) in FACS buffer.

Techniques: Derivative Assay, Incubation, Recombinant, Saline, Expressing, Flow Cytometry, Real-time Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Tereticornate A suppresses RANKL-induced osteoclastogenesis via the downregulation of c-Src and TRAF6 and the inhibition of RANK signaling pathways.

doi: 10.1016/j.biopha.2022.113140

Figure Lengend Snippet: Fig. 3. TA inhibits osteoclast-specific protein and gene expression during osteoclastogenesis. (A,B) Representative immunoblots for the effects of TA on RANKL- induced expression of TRAP (A) and cathepsin K (B) proteins. The gray densities of the bands for TRAP and cathepsin K were quantified using the AlphaView software. (C–F) TA inhibited RANKL-induced mRNA expression of β-Integrin (C), MMP-9 (D), ATP6V0D2 (E), and DC-STAMP (F). Total proteins or RNA from RAW 264.7 cells pretreated with or without TA for 15 min, then stimulated with RANKL (50 ng/mL) for 24 h, were subjected to western blotting or qRT-PCR analysis. Data are presented as the mean ± SEM from three independent biological experiments. ***p < 0.001 compared to the control group; #p < 0.05, ##p < 0.01, and ###p <

Article Snippet: Primary antibodies against NFATc1, c-Fos, TRAP, cathepsin K, TRAF6, and c-Src were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Gene Expression, Western Blot, Expressing, Software, Quantitative RT-PCR, Control

Journal: Scientific reports

Article Title: Disordered metabolism in mice lacking irisin.

doi: 10.1038/s41598-020-74588-7

Figure Lengend Snippet: Figure 6. Increased bone resorption in irisin lacking mice. (A, B) Sections of the distal shaft of the femur were stained with osteoprotegerin (OPG) (1:60) and receptor activator of nuclear factor-kB ligand (RANKL) (1:80) antibodies. Positive (black arrows) osteoblasts are shown, 200×. (C) Representative images of the distal metaphyseal region of the femur, together with cell counts (osteoclasts) per bone perimeter (Bpm). Red arrows, tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts, 100×, (NIS-Elements Viewer, v4.2.0; https:// www.downza.cn/soft/2751-21.html); (D) serum levels of osteocalcin TRAP (with three replicates). (Graphpad Prism, v7.0, https://www.xue51.com/soft/3932.html). Data are presented as the mean ± SEM (n = 15 per group). *P < 0.05, **P < 0.01 compared to the WT group.

Article Snippet: The enzyme-linked immunosorbent assay (ELISA) kit for osteocalcin (OCN) (E06917m), Alkaline Phosphatase (ALP) (E0200m), and Tartrate-resistant acid phosphatase (TRAP) (E08492m) were purchased from Elabscience (Wuhan, China).

Techniques: Staining