Journal: Microcirculation (New York, N.y. : 1994)
Article Title: Regulatory T Cells Control Vascular Adhesion Molecule Expression in Skin Under Inflammatory and Homeostatic Conditions
doi: 10.1111/micc.70017
Figure Lengend Snippet: CHS induces upregulation of E‐selectin and ICAM‐1 in the dermal microvasculature. Adhesion molecule expression in the dermal microvasculature in a two‐challenge model of CHS was assessed in wild‐type (WT) C57BL/6 mice 4 h after the second challenge, via in vivo labelling with fluorochrome‐conjugated antibodies. Tissues underwent intravascular staining followed by exsanguination, fixation, clearing and imaging via multiphoton microscopy, with samples collected from CHS skin and untreated skin from the same mice. (A, B) Representative images of staining for CD31 (vasculature), E‐selectin and ICAM‐1 (A) or CD31, P‐selectin and VCAM‐1 (B) either in untreated skin (top row in A, B) or during CHS (bottom row in A, B). Images are maximum intensity projections through the skin, taken from the epithelial surface to ~400 μm depth in inflamed samples. Right‐hand panels show merged images. Scale bars = either 200 μm (A: WT untreated; B: WT CHS) or 150 μm (A: WT CHS; (B) WT Untreated). (C) Isotype control antibody staining in the dermal microvasculature during two challenge CHS. Mice underwent the two‐challenge model of CHS and received AF594‐anti‐CD31, along with AF680‐mouse IgG1κ (isotype control for RME‐1/anti‐E‐selectin) and AF488‐rat IgG2b (isotype control for YN1.1.7.4/anti‐ICAM‐1), and the skin was cleared and imaged as for the other samples. Images are shown for staining for CD31 (red), and the AF680‐conjugated (cyan) and AF488‐conjugated (green) isotype control antibodies. Scale bar = 150 μm. (D, E) Quantitation of adhesion molecule expression in untreated (UT) or CHS skin for E‐selectin and P‐selectin (D) or ICAM‐1 and VCAM‐1 (E). Data from samples treated with isotype control antibodies conjugated with AF680 (D) or AF488 (E) are also shown. Data are shown as mean ± sem with data for individual mice also shown. N = 3/group for adhesion molecules and 4/group for isotype controls. * p < 0.05 for comparison of UT and CHS (in the same animal) via paired t ‐test for each adhesion molecule.
Article Snippet: To enable visualization of the vasculature, AF680‐albumin (25 μg/mouse, Thermo Fisher Scientific) was administered i.v.
Techniques: Expressing, In Vivo, Staining, Imaging, Microscopy, Control, Quantitation Assay, Comparison