Journal: International journal of molecular sciences
Article Title: Hematopoietic Growth Factors Regulate the Entry of Monocytes into the Adult Brain via Chemokine Receptor CCR5.
doi: 10.3390/ijms25168898
Figure Lengend Snippet: Figure 1. Receptors for SCF and G-CSF are expressed on brain endothelial cells. (A,B) Representative 3-dimensional (3D) confocal images. Capillaries (CD31+, red) in the cortex of the adult mouse brain show co-expression with the receptors for SCF (A) (Ckit, green) and G-CSF (B) (GCSFR, green). Scale bar, 10 µm. (C,D) Representative flow cytometry data. The expressions of Ckit (C) and GCSFR (D) on cerebral endothelial cells are detected by flow cytometry analysis of CD45−CD31+ gated endothelial cells isolated from adult mouse brain. Mean fluorescence intensity (MFI) of Ckit (C) and GCSFR (D) expressing on cerebral endothelial cells is presented in flow cytometry histograms. Isotype: isotype control antibody. Repeated experiments, n = 4. (E,F) Representative confocal images of immunofluorescence staining. The expression of Ckit (E) (red) and GCSFR (F) (red) are seen on bEnd.3 endothelial cells (CD31+, green). Scale bar, 50 µm. Repeated experiments, n = 3. (G,H) Representative flow cytometry histograms showing the expression of Ckit (G) and GCSFR (H) on bEnd.3 endothelial cells. MFI: Mean fluorescence intensity. Isotype: isotype control antibody. Repeated experiments, n = 5.
Article Snippet: Anti-mouse antibodies used for flow cytometry included: rat anti-CD31 (MEC 13.3, BD Pharmingen, San Diego, CA, USA), rat anti-CD45-APC (30F11, eBioscience, San Diego, CA, USA), rabbit anti-mouse ckit (sc-168, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-mouse GCSFR (sc-694, Santa Cruz Biotechnology, USA), rabbit IgG (sc-3888, Santa Cruz Biotechnology, USA), anti-CCR5-PE (HM-CCR5 (7A4), eBioscience, USA), antiCD106-FITC (VCAM-1, 429, eBioscience, USA), and anti-CD54-FITC (ICAM-1, YN1/1.7.4, eBioscience, USA).
Techniques: Expressing, Flow Cytometry, Isolation, Fluorescence, Control, Immunofluorescence, Staining
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