Journal: Nature Communications

Article Title: Single-cell reconstruction of follicular remodeling in the human adult ovary

doi: 10.1038/s41467-019-11036-9

Figure Lengend Snippet: Complement and immune system in the adult ovaries. a tSNE cluster map showing expression of selected complement genes. b Gene network of C1S in TC and stroma. The color of the circles represents function and the color of the edges represent networks. c Immunostaining of follicles (ø, diameter) growing (top row), atretic (middle row), and degenerated (bottom row) for C1S, C1Q, and CD68. Single channel images were converted to an intensity map. Scale bars are 100 μm. d Immunostaining of follicles (ø, diameter) growing (top row), atretic (middle row), and degenerated (bottom row) for C1Q and STAR. Scale bars are 100 μm. e Concentration of secreted C1Q and C3 produced by pieces of human ovarian stroma ( N = 3 samples) and atretic follicle walls ( N = 6 samples) after 1 and 5 days of culture. Median and sample distribution (dots) are shown. f tSNE cluster map showing expression of C1QA and C3 . g–i tSNE cluster map showing expression of selected immune marker genes for innate immune cells g , B lymphocytes h and T lymphocytes and NK cells i . Red dashed lines give the boundaries of the specific immune-clusters

Article Snippet: The primary antibodies used were rabbit anti-KI67 (1:100, ab15580, Abcam), mouse anti-AMH (1:30, MCA2246T, BioRad), rabbit anti-Troponin I (H-170) (1:100, sc15368, Santa Cruz), goat anti-VASA/DDX4 (1:200, AF2030, R&D), mouse anti-Cytokeratin (1:100, M351501, DAKO), rabbit anti-Wilm’s Tumor protein (1:100, CA1026-50, Calbiochem), mouse anti-StAR (D-2) (1:100, sc166821, Santa Cruz), rabbit anti-alpha smooth muscle actin/ACTA2 (1:200; ab5694, Abcam), goat anti-ZP3 (N-20) (1:100, sc23715, Santa Cruz), mouse anti-Connexin-43/GJA1 (CX-1B1) (1:50, 13-8300, Zymed), rabbit anti-Fragilis/IFITM3 (1:200, ab15592, Abcam), rabbit anti-Collagen Type IV (1:50, AB748, Chemicon), rabbit anti-Von Willebrand factor/VWF (1:100, ab6994, Abcam), mouse anti-N-Cadherin/CDH2 (GC-4) (1:100, C3865, Sigma-Aldrich), rabbit anti-PECAM1 (M-20) (1:200, sc1506, Santa Cruz), rabbit anti-c-FOS (1:20, PC38, Calbiochem), mouse anti-8-OHdG (1:1000, sc66036, Santa Cruz), goat anti-IGFBP5 (1:50, AF875, R&D), rabbit anti-C1Q (1:400, A0136, DAKO), goat anti-C1S (1:400, A302, Quidel) and mouse anti-CD68 (1:50, M087629-2, DAKO).

Techniques: Expressing, Immunostaining, Concentration Assay, Produced, Marker

Journal: Frontiers in Immunology

Article Title: Role of CCR2 + Myeloid Cells in Inflammation Responses Driven by Expression of a Surfactant Protein-C Mutant in the Alveolar Epithelium

doi: 10.3389/fimmu.2021.665818

Figure Lengend Snippet: Effects of CCR2 monocyte ablation on inflammatory and fibrotic pathways following SP-C I73T induced injury. Histochemical analysis of control (CTL, tamoxifen treated SP-C WT or oil treated SP-C I73T mice), SP-C I73T CCR2 WT and SP-C I73T CCR2 KO lung sections 14 d post injury immunostained with antibody to (A) C1q and (B) p-SMAD2/3. Binding was visualized using a Vectastain kit. Red arrowheads indicate macrophage expression; grey arrowheads indicate non-macrophage expression. Insets show magnified cells; box color matches that of arrowhead. Original magnification, 400x; inset magnification, 750x. Representative sections from 3 mice/group are shown.

Article Snippet: Antibodies used for in situ hybridization, immunohistochemical and flow cytometric analysis were: tgfb1 (Cat # 407751, Advanced Cell Diagnostics, ACD); RFP (Cat # ab62341; 1:1000, Abcam, Cambridge, MA); Arg1 (Cat # ab91279; 1:1500, Abcam); iNOS (Cat # ab15323; 1:200, Abcam); CD64 (Cat # bs-3511R; 1:250, Bioss Antibodies, Woburn, MA); CCR2 (Cat # EPR20844; 1:400, Abcam); phospho-SMAD2/3 (Cat # PA5-37636; 1.500, Invitrogen CX 3 CR1 (Cat # bs-1728R; Bioss Antibodies); CD64 (Cat # bs-3511R; Bioss Antibodies); CD125/IL-5RA (Cat # bs-2601R, Bioss Antibodies); C1q (Cat # A0136; 1:500, Dako/Agilent Technologies, Santa Clara, CA); CD16/32 (clone 93; eBiosciences, San Diego, CA), CD11b (clone # M1/70; eFluo450, eBiosciences); Fixable Viability dye (Cat # 65-0865-14; eFluo780, eBiosciences); SigF (clone S17007L; PE-CF594, BD Biosciences, San Jose, CA); CD45 (clone 30-F11; PerCP5.5, Biolegend, San Diego, CA); CD11c (clone # N418; BV705, Biolegend); Ly6G (clone # 1A8; AF700, Biolegend); Ly6C (clone HK1.4, BV510, Biolegend); CD64 (clone X54-5/7.1; PE/Cy7, Biolegend); CD43 (clone # S11; PE, Biolegend); CD3 (clone # 17A2; BUV395, Biolegend).

Techniques: Binding Assay, Expressing