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

Journal: Cancer Immunology Research

Article Title: Tumor-Associated Neutrophils Drive B-cell Recruitment and Their Differentiation to Plasma Cells

doi: 10.1158/2326-6066.cir-20-0839

Figure Lengend Snippet: Figure 5. TANs drive B-cell differentiation independently of T cells. A, Quantification of CD138 expression on splenic B220þ cells following various culture conditions—when cultured alone (B), with T cells (B þ T, ratio 1:1), with TANs (B þ TAN, ratio 1:1), with TAN and T cells (B þ TAN þ T, ratio 1:1:1), with T cells but no contact allowed with TANs (B þT//TAN,ratio 1:1:1), or TANs cocultured with T cells but no contact allowed with B cells (B//T þTAN, ratio 1:1:1). Data represent the mean SEM (n ¼ 6–9; , P < 0.001; n.s., not significant). Groups were compared using one-way ANOVA. B, Representative flow plots displaying CD138 expression out of total B220þ population in the different coculture conditions. C, Quantification of IgG release to the media following overnight coculture of isolated splenic B220þ

Article Snippet: Isotype control antibodies were as follows: APCconjugated rat IgG2a (clone 2A3; Biogems), FITC-conjugated rat IgG2b (clone RTK4530; BioLegend), BGViolet450 rat IgG2a (clone 2A3, Biogems), PE-conjugated rat IgG2a (clone 2A3, Biogems).

Techniques: Cell Differentiation, Expressing, Cell Culture, Isolation

Journal: Cancer Immunology Research

Article Title: Tumor-Associated Neutrophils Drive B-cell Recruitment and Their Differentiation to Plasma Cells

doi: 10.1158/2326-6066.cir-20-0839

Figure Lengend Snippet: Figure 6. TANs express membranal BAFF, but not membranal APRIL, and mediate B-cell IgG production in a BAFF-R–dependent manner. A and B, Expression of membranal BAFF and APRIL in TANs within the whole tumor (A) and following TANs' isolation from the tumors (B). Representative histograms showing BAFF and APRIL expression in TANs (gated as total Ly6Gþ population) are provided (right plots). C, IgG production by splenic B cells cocultured with TANs (ratio 1:5) in the absence or presence of anti–BAFF-R antibody. Data represent the mean SEM (n ¼ 4; , P < 0.01; , P < 0.001). Groups were compared using one-way ANOVA. D, Quantification of CD138 expression on isolated splenic B220þ cells cultured alone or cocultured with TANs (ratio 1:5), without or with blocking of the three potential BAFF receptors, BAFF-R, TACI, or BCMA. Data represent the mean SEM (n ¼ 4–10; , P < 0.001; n.s., nonsignificant). Groups were compared using one-way ANOVA.

Article Snippet: Isotype control antibodies were as follows: APCconjugated rat IgG2a (clone 2A3; Biogems), FITC-conjugated rat IgG2b (clone RTK4530; BioLegend), BGViolet450 rat IgG2a (clone 2A3, Biogems), PE-conjugated rat IgG2a (clone 2A3, Biogems).

Techniques: Expressing, Isolation, Cell Culture, Blocking Assay