Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: Anti-IL-9 antibody treatment decreased mast cell infiltration of the CNS. Purified cells (mainly composed of mast cells) were harvested from the CNS on days 0, and 5, 15, and 20 after EAE immunization. CD45 + CD117 + mast cells were detected by flow cytometry. CNS mast cells maintained a high level throughout the disease course in EAE group. However, mast cells were significantly reduced from day 5, and remained low in anti-IL-9 Abs group, compared with IgG group. Data are shown as mean ± SE. * P < 0.01, anti-IL-9 Abs group versus IgG group. CNS: Central nervous system; EAE: Experimental autoimmune encephalomyelitis; SE: Standard error; IL: Interleukin.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: Purification, Flow Cytometry

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: IL-9 blockade reduced production of chemokine recruiting mast cells in the CNS. Five days after immunization, mRNA expressions of Pecam1, SCF, Vcam-1, CCL2, and CCL5 in CNS tissue of EAE mice were detected by RT-PCR. After IL-9 neutralization, mRNA expressions of CCL5 and Vcam-1 were significantly decreased in anti-IL-9 Abs group, compared with IgG group. * P < 0.01. Pecam1: Platelet and endothelial cell adhesion molecule 1; SCF: Supercoiling factor; Vcam-1: Vascular cell adhesion molecule 1; CCL2: C-C motif chemokine ligand 2; CCL5: C-C motif chemokine ligand 5; CNS: Central nervous system; EAE: Experimental autoimmune encephalomyelitis; IL: Interleukin; RT-PCR: Reverse transcription-polymerase chain reaction; mRNA: Messenger RNA.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Neutralization

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: In vitro , the effect of anti-IL-9 antibody on splenic mast cells. Splenocytes were harvested from experimental autoimmune encephalomyelitis mice 5 days after MOG immunization. After co-culture with anti-IL-9 antibody or anti-mouse IgG for 7 h, mast cell number was counted by flow cytometry. Splenic mast cells cultured with anti-IL-9 antibody showed significantly lower levels in a dose-dependent manner. This trend was particularly evident with anti-IL-9 antibody concentrations up to 20 μg/ml. * P < 0.05; † P < 0.01. IL: Interleukin; MOG: Myelin oligodendrocyte glycoprotein.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: In Vitro, Co-Culture Assay, Flow Cytometry, Cell Culture

Journal: iScience

Article Title: Microglial transcription profiles in mouse and human are driven by APOE4 and sex

doi: 10.1016/j.isci.2021.103238

Figure Lengend Snippet:

Article Snippet: Cells were then rinsed in PBS, and permeabilized using 0.1% Triton X-100 in PBS for 5 min. After blocking in 2% BSA for 1h, cells were incubated overnight at 4°C in primary antibodies directed against IBA1 (1:500, Wako), CD11c (1:500, Novus Biologicals), purinergic receptor P2Y (P2ry12; #HPA013796, 1:500, Atlas Antibodies), lysozyme C (LYZ; #sc27958, 1:100, Santa Cruz Biotechnology), endoglin (CD105; #MAB1097, 1:100, R&D Systems), or cleaved caspase 3 (CC3; #9664, 1:1000, Cell Signaling Technology).

Techniques: Plasmid Preparation, Recombinant, Gene Expression, Generated, Software

Journal: Journal of pharmacological sciences

Article Title: Oxymatrine alleviates NSAID-associated small bowel mucosal injury by regulating MIP-1/CCR1 signalling and gut microbiota.

doi: 10.1016/j.jphs.2024.08.003

Figure Lengend Snippet: Fig. 4. OMT alleviated NSAID-associated small intestinal mucosal barrier disorder. (A) IHC analysis of the protein expression of Zo-1 and Occludin in the ileum tissues from the four groups. (B) IHC analysis of the protein expression of LYZ and CHGA in the ileum tissues from the four groups. (C) IHC analysis of the protein expression of MUC2 in the ileum tissues from the four groups.*P < 0.05 vs. the control group, #P < 0.05 vs. DS group.

Article Snippet: The primary antibodies against CHGA (ab283265, Abcam, United Kingdom), LYZ (66456-i-Ig, Proteintech, China), MUC2 (ab272692, Abcam, United Kingdom), Ki-67 (ab16667, Abcam, United Kingdom), myc (Ab32072, Abcam, United Kingdom), caspase3 (GB11009-1, Servicebio, China), CD68 (GB113109, Servicebio, China), IL-6 (GB11117, Servicebio, China), IL-1β (GB11113, Servicebio, China), TNF alpha (60291-i-Ig, Proteintech, China), Zo-1 (21773- 1-AP, Proteintech, China), Occludin (ab216327,Abcam, United Kingdom), MIP-1γ (GB112361, Servicebio, China), CCR1 (A18341, Abclonal, China), P38 MAPK (A14401, Abclonal, China), phospho-p38 MAPK (AF4001, Affinity), phospho–NF–KB p65 (AP0944, Abclonal), NF-KB p65 (8242,CST), MIP-1α (bs-1045r, BIOSS), CCR1 (bs-1169r, BIOSS) and GAPDH (GB15004, Servicebio) were used in the follow-up study.

Techniques: Expressing, Control