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: bioRxiv

Article Title: Macrophages regulate meiotic initiation and germ cell clearance in the developing ovary

doi: 10.64898/2026.02.28.708733

Figure Lengend Snippet: (a) Representative whole-mount images of E13.5 ovaries from C57BL/6J embryos exposed at E6.5 to either control rat IgG2a antibody or anti-CSF1R blocking antibody to deplete YS-derived macrophages. For F4/80, PECAM1, and FOXL2 staining, n =5 control and n =6 anti-CSF1R-treated independent gonads; for CD11b and CD45 staining, n =3 control and n =3 anti-CSF1R-treated independent gonads. Insets show higher-magnification views of boxed regions; dashed lines mark the gonad-mesonephros boundary. (b) qRT-PCR analysis of macrophage-associated genes ( Adgre1, Cx3cr1, Csf1r, Mrc1 ) and endothelial marker Cdh5 , showing fold change in gene expression in E13.5 anti-CSF1R-treated ovaries versus controls. (c) Representative E14.5 sections stained for SYCP3 and TRA98 showing increased meiotic germ cells in anti-CSF1R-treated ovaries ( n =4) compared with control ovaries ( n =4). (d) qRT-PCR analysis of germ cell and meiotic genes ( Kit, Pou5f1, Ddx4, Stra8, Sycp1, Sycp3, Syce1, Smc1b ), showing fold change in gene expression in E14.5 anti-CSF1R-treated ovaries versus controls. (e) Representative E18.5 sections stained for F4/80 and CD45, and for SYCP3 and FOXL2, showing recovery of F4/80⁺ macrophages and advanced meiotic progression in anti-CSF1R-treated ovaries compared with control ovaries. For F4/80 and CD45 staining, n =4 control and n =4 anti-CSF1R-treated independent gonads; for SYCP3 and FOXL2 staining, n =3 control and n =3 anti-CSF1R-treated independent gonads. Arrows indicate SYCP3⁺ dictyate-stage oocytes. (f) Percentage of SYCP3⁺ germ cells at the dictyate stage among total SYCP3⁺ cells at E18.5. Data are shown as mean +/− SD. * P <0.05, ** P <0.01, *** P <0.001 (two-tailed Student’s t -tests). Scale bars, 100 μm (overview) and 50 μm (higher magnification/insets).

Article Snippet: To transiently deplete yolk-sac-derived and fetal CSF1R + macrophages, pregnant C57BL/6J females were injected intraperitoneally with 3 mg anti-CSF1R monoclonal antibody (mAb; clone AFS98, Bio X Cell #BP0213) or rat IgG2a isotype control (Bio X Cell #BP0089).

Techniques: Control, Blocking Assay, Derivative Assay, Staining, Quantitative RT-PCR, Marker, Gene Expression, Two Tailed Test

Journal: bioRxiv

Article Title: Macrophages regulate meiotic initiation and germ cell clearance in the developing ovary

doi: 10.64898/2026.02.28.708733

Figure Lengend Snippet: (a) Representative images of E18.5, P3, and P10 ovaries from C57BL/6J embryos exposed at E6.5 and E14.5 to either control rat IgG2a antibody or anti-CSF1R blocking antibody to deplete fetal ovarian macrophages. For E18.5, n =4 control and n =4 anti-CSF1R-treated independent gonads; for P3, n =6 control and n =6 anti-CSF1R-treated independent gonads; for P10, n =4 control and n =4 anti-CSF1R-treated independent gonads. Insets show higher-magnification views of boxed regions; red and green dashed outlines demarcate the ovary. (b) Representative sections stained for DDX4, showing developmental germ cell attrition at E18.5, P3, and P10 in control and anti-CSF1R-treated ovaries. For each time point, n =4 control and n =4 anti-CSF1R-treated independent gonads. (c) Quantification of CD45⁺IBA1 + macrophages per 0.1 mm² ovarian area at E18.5, P3, and P10. (d) Quantification of CD45⁺IBA1 − monocyte-like cells per 0.1 mm² ovarian area at the indicated stages. (e) Quantification of DDX4⁺ germ cell number per 0.1 mm² ovarian area at E18.5, P3, and P10, showing reduced physiological germ cell loss after macrophage depletion. Data are shown as mean +/− SD. * P <0.05, ** P <0.01, *** P <0.001 (two-tailed Student’s t -tests). Scale bars, 100 μm (overview) and 50 μm (higher magnification/insets).

Article Snippet: To transiently deplete yolk-sac-derived and fetal CSF1R + macrophages, pregnant C57BL/6J females were injected intraperitoneally with 3 mg anti-CSF1R monoclonal antibody (mAb; clone AFS98, Bio X Cell #BP0213) or rat IgG2a isotype control (Bio X Cell #BP0089).

Techniques: Control, Blocking Assay, Staining, Two Tailed Test