marco  (R&D Systems)

Journal: JCI Insight

Article Title: Csk-mediated Src family kinase regulation dampens neutrophil infiltration during pulmonary infection

doi: 10.1172/jci.insight.188323

Figure Lengend Snippet: ( A – K ) Csk fl/fl Lyz2 wt/wt and Csk fl/fl Lyz2 cre/wt mice were subjected to K . pneumoniae intratracheal injection or sham surgery. After 24 hours, bacterial burden regarding the colony forming units (CFUs) in lung ( A ), bronchoalveolar fluid (BALF) ( B ), blood ( C ), and spleen ( D ) and neutrophil recruitment into the lungs ( E ) and BALF ( F ) were determined. ( G ) Neutrophil recruitment (CD45 + Gr-1 + LyB.2 + ) in Csk fl/fl Lyz2 wt/wt and Csk fl/fl Lyz2 cre/wt mice in the interstitial, intravascular, and BALF compartments following intratracheal instillation of K . pneumoniae . Cell count of monocytes (MO; CD45 + CD11b + CX3CR1 + Ly6C hi Ly6G – Gr-1 – ) ( H ), alveolar macrophages (AM; CD45 + CD64 + F4/80 + MARCO + SiglecF hi ) ( I ), natural killer cells (NK; CD45 + CD27 + CD335 + ) ( J ), and dendritic cells (DC; CD45 + CD27 – CD24 + CD11c + MHCII + ) ( K ) in lung tissue 24 hours after lung infection. n as indicated, mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 by 1-way-ANOVA by Holm-Šídák multiple-comparison test ( A – D ), 1-way ANOVA with Tukey’s multiple-comparison test ( E , F , and H – K ), or 2-tailed Student’s t test ( G ).

Article Snippet: Alveolar macrophages were gated as CD45 + CD64 + F4/80 + MARCO + SiglecF hi using antibodies against CD45 (clone 30-F11, BioLegend), CD64 (clone X54-5/7.1, BioLegend), F4/80 (clone BM8, BioLegend), MARCO (clone 579511, R&D Systems), and SiglecF (clone S17007L, BioLegend).

Techniques: Injection, Cell Counting, Infection, Comparison

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: AXL promotes lymphangiogenesis by amplifying VEGF-C-mediated AKT pathway

doi: 10.1007/s00018-024-05542-3

Figure Lengend Snippet: AXL is expressed by MARCO + LECs in murine LN. A. Representative images of naive murine lymph node immunostained for AXL (red), LYVE-1 (green), MARCO (white). B-D . In vivo Tumor-associated lymphangiogenesis assay. The experimental design of the assay is given in ( B ). Gelatin sponges containing (tumor draining: Td) or not (sham) melanoma cells were implanted in mice ear. The sentinel LN (Td LN or sham LN) were resected 28 days later. Images of AXL (red), MARCO (white), LYVE-1 (green) and DAPI (nuclei, blue) were acquired ( C ). Computerized quantification of the staining densities ( D ) and spatial distribution analysis ( E , F ) were performed in murine Sham and Td LNs. D. Data are means ± SEM for LYVE-1 and MARCO graphs and medians with interquartile ranges for AXL graph. Statistical tests: Unpaired t test (LYVE-1 and MARCO graphs) and Mann–Whitney U test (AXL graph). * p < 0.05, ** p < 0.01. E. Data are staining area medians according to the distance from the LN border (d = 0). F. Data are p-values of staining area differences between Td and Sham LNs for each distance. Below the dotted line, p is lower than 0.05 and considered as significant. Statistical test: Mann–Whitney U test for each distance. n ≥ 7 for each group. G. Images of Prox1 RNAscope hybridization (lymphatic vessel, green) and AXL immunostaining (red) in Sham and Td LNs

Article Snippet: Cryosections were incubated at room temperature for 1 h with AXL and MARCO (Bio-Rad Cat# MCA1849T, RRID: AB_2140591, 1/200) antibodies.

Techniques: In Vivo, Staining, MANN-WHITNEY, RNAscope, Hybridization, Immunostaining