Mast cell chymase

Rabbit anti-ICAM-1, VCAM-1, PECAM-1 and GAPDH polyclonal antibodies; rat anti- DC Marker (33D1); and mouse anti-SRA, αSMA, CD36, Arg1, SRBI, LOX-1, NK Cell Marker, CD4, CD22, Mast Cell Chymase and CD68 monoclonal antibody were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Rabbit anti-ABCG1, ABCA1 and Asialo GM1 polyclonal antibodies were purchased from Novus Biologicals (Littleton, CO). Rabbit anti-iNOS polyclonal antibodies were purchased from Proteintech Group, Inc. (Rosemont, IL). Rabbit anti-KLF4 antibody and anti-Lamin A/C monoclonal antibody were purchased from Abcam (Cambridge, MA). Mouse anti-rabbit IgG-R, mouse anti-rabbit IgG-FITC and m-IgGκ BP-FITC antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Formononetin was purchased from Yuanye (Shanghai, China).

Immunofluorescence staining of GFAP, IBA1, Mast Cell Chymase, Mast Cell Tryptase, CD4, and CD8 was conducted as previously indicated [50 (link)]. Following deparaffinization, the following primary antibodies were incubated overnight on the brain slices.: GFAP (1:100; sc-9065 Santa Cruz Biotechnology, Santa Cruz, CA, USA), IBA-1 (1:100; sc-32725 Santa Cruz Biotechnology, Santa Cruz, CA, USA), Mast Cell Chymase (1:100; sc-59586 Santa Cruz Biotechnology, Santa Cruz, CA, USA), Mast Cell Tryptase (1:100; sc-59587 Santa Cruz Biotechnology, Santa Cruz, CA, USA), CD-4 (1:100; sc-514571 Santa Cruz Biotechnology, Santa Cruz, CA, USA), and CD-8 (1:100; sc-7970 Santa Cruz Biotechnology, Santa Cruz, CA, USA). The secondary fluorescent antibodies Alexa Flour 488 goat anti-mouse (A11001; Molecular Probes, Eugene, OR, USA) or anti-rabbit IgG (A11008; Molecular Probes, Eugene, OR, USA) were incubated for three hours the next day, depending on the primary antibody employed. Subsequently, slices were stained with 4′,6′-diamidino-2-phenylindole (DAPI; Hoechst, Frankfurt, Germany). The histological studies were carried out in a blinded manner, and a fluorescence microscope picture (Nikon Eclipse Ci-L microscope, NIKON CORPORATION, Tokyo, Japan) was used to collect. Pictures are shown at 40× magnification.