Journal: The Journal of investigative dermatology

Article Title: Overexpression of cathepsin S induces chronic atopic dermatitis in mice.

doi: 10.1038/jid.2011.404

Figure Lengend Snippet: Figure 4. Proteinase-activated receptor-2 (PAR-2) expression and inflammatory cell infiltration in dorsal skin. Real-time PCR analysis of (a) cathepsin S (CTSS) and (b) PAR-2 expression. Diamonds, C57BL/6 WT mice; squares and triangles, CTSS-overexpressing TG mice (n ¼ 4). The average value for WT mice was set to 1. *Po0.05, diamonds and squares; #Po0.05, ##Po0.01, squares and triangles. (c) Western blot analysis of dorsal skin tissue homogenates with antibodies specific for CTSS, PAR-2, and b-actin (arrow). Immunostaining of dorsal skin with antibodies specific for (d) CTSS, PAR-2, and CD11c, as well as (e) major histocompatibility complex (MHC) class II, CD4 (CD4 þ T-cell marker), and CD68 (macrophage marker). Markers of mature dendritic cells: CD11c and MHC class II. Bar ¼ 50 mm. Positive cells show brown staining. TG, CTSS-overexpressing transgenic mice; WT, C57BL/6 wild-type mice.

Article Snippet: The following antibodies were used: anti-mouse cytokeratin 10 (K10) monoclonal antibody (Covance, Princeton, NJ), anti-rabbit Ki67 monoclonal antibody (Thermo, Hudson, CA), anti-mouse proliferat- ing cell nuclear antigen monoclonal antibody, anti-goat CTSS poly- clonal antibody, anti-rabbit PAR-2 polyclonal antibody, anti-mouse MHC class II monoclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA), anti-hamster CD11c monoclonal antibody, anti-rabbit CD4 polyclonal antibody (Novus Biologicals, Littleton, CO), and anti-rat macrophage monoclonal antibody (Abcam, Cambridge, UK).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunostaining, Immunopeptidomics, Marker, Staining, Transgenic Assay

Journal: STAR Protocols

Article Title: Protocol to isolate immune cells from mouse pancreatic lymph nodes and whole pancreas for mass cytometric analyses

doi: 10.1016/j.xpro.2022.101938

Figure Lengend Snippet: Examples of manual gating for different immune cells within the CD45+ population from the pancreatic lymph node of 8-week-old female NOD mice (A) Example of gating for monocytes (CD11b+). (B) Example of gating for dendritic cells (CD11c+). (C) Example of gating for B cells (CD19+). (D) Example of gating for CD4+ T cells (CD4+). (E) Example of gating for CD8+ T cells (CD8+). (F) Example of gating for macrophages (F4/80+).

Article Snippet: Anti-mouse CD11c, Clone N418, 1:50 dilution , Fluidigm , Cat#3142003C, Label 142Nd.

Techniques:

Journal: STAR Protocols

Article Title: Protocol to isolate immune cells from mouse pancreatic lymph nodes and whole pancreas for mass cytometric analyses

doi: 10.1016/j.xpro.2022.101938

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD11c, Clone N418, 1:50 dilution , Fluidigm , Cat#3142003C, Label 142Nd.

Techniques: Recombinant, Staining, Red Blood Cell Lysis, Software, Cytometry

Journal: Frontiers in Pharmacology

Article Title: Protective Effects of Live Combined B. subtilis and E. faecium in Polymicrobial Sepsis Through Modulating Activation and Transformation of Macrophages and Mast Cells

doi: 10.3389/fphar.2018.01506

Figure Lengend Snippet: The effect of LCBE pretreatment on the macrophage activation and transformation. (A) CD68was stained in ileal tissues by IHC assay under 40× and 100× magnifications. (B) The protein expression of CD11cwas examined by Western blotting. Representative photographs of CD11c and β-actin blots were displayed in left panel. The bar graph was presented in right panel. (C) The protein expression of M2 type macrophage marker CD206 was examined. Representative photographs of CD206 and β-actin blots were exhibited in left panel. The bar graph was presented in right panel. All data were showed as means ± SEM. N = 6 mice/group; N = 6 mice/group; ∗ P < 0.05.

Article Snippet: Samples containing equal amounts of protein (30 μg) per lane were fractionated based on their molecular weight by 8–10% sodium dodecyl sulfate-polyacrylamide gels for electrophoresis (SDS-PAGE) and transferred onto polyvinylidene difluoride (PVDF) membranes at a constant current of 200 mA for 60 min. After blocking with 5% non-fat milk in Tris-buffered saline with 0.1% Tween-20 (TBST) for 2 h at room temperature, the membranes were incubated with primary antibodies against β-actin (1:5000, as a reference control; Abmart, United States), TGF-β1 (1:1000; Abcam, United States), CD11c (1:500; Proteintech, United States), CD206 (1:500; Proteintech, United States), p-AKT (1:1500; Abcam, United States), Akt (1:2000; Abcam, United States), Claudin (1:500; Sigma, Shanghai), Occludin (1:1000, Sigma, Shanghai), and ZO-1 (1:500, Sigma, Shanghai) overnight at 4°C.

Techniques: Activation Assay, Transformation Assay, Staining, Expressing, Western Blot, Marker