Journal: Nature

Article Title: The transition from monocyte to tissue-resident macrophage requires DHPS

doi: 10.1038/s41586-025-09972-2

Figure Lengend Snippet: Representative immunofluorescence images of a, liver, and b, spleen from adult Dhps +/+ Rosa26 eYFP and Dhps -ΔM Rosa26 eYFP mice. Total macrophages were stained with fluorescently-tagged antibodies against F4/80 for liver and spleen sections, and TIM-4 to detect Kupffer cells in liver sections and TIM-4 expressing cells in spleen. Anti-GFP was used to detect YFP + cells (LysM Cre), and nuclei were visualized by DAPI staining. Quantification of F4/80 + , F4/80 + YFP + and F4/80 + YFP + TIM-4 + cells per unit area (mm 2 ) in Dhps +/+ Rosa26 eYFP and Dhps -ΔM Rosa26 eYFP mice is shown for each tissue. The percentage of TIM-4 + cells within the total F4/80 + YFP + population was calculated for both tissues. All images and graphs are representative of 4–5 mice per group. Quantification for liver was performed across the entire tissue section and expressed as cells per unit area. Data are mean ± s.d. Exact P-values are indicated. Red boxes indicate RTM populations. Illustrations in a and b were created using BioRender ( https://biorender.com ).

Article Snippet: Slides were cover-slipped with EverBrite TrueBlack Hardset Mounting Medium with DAPI (Biotium, 23018) or ProLong Gold Antifade Mountant (Invitrogen, P36930 ) after nuclear staining with 1 μg ml −1 DAPI (Thermo Scientific, 62248) for 10 min at room temperature.

Techniques: Immunofluorescence, Staining, Expressing

Journal: Nature

Article Title: The transition from monocyte to tissue-resident macrophage requires DHPS

doi: 10.1038/s41586-025-09972-2

Figure Lengend Snippet: a , Representative immunofluorescence images of kidney from adult Dhps +/+ CX3CR1-ERT2 Cre Rosa26 eYFP (control) and Dhps flox/flox CX3CR1-ERT2 Cre Rosa26 eYFP mice 5 weeks after in vivo tamoxifen administration. CX3CR1-ERT2 Cre-targeted kidney resident macrophages were stained with fluorescently-tagged antibodies against GFP to detect YFP + cells (CX3CR1-ERT2Cre). Nuclei were visualized by DAPI staining. b , Quantification of YFP + kidney macrophages per unit area (mm 2 ) in Dhps +/+ (control) and Dhps flox/flox CX3CR1-ERT2 Cre Rosa26 eYFP mice are shown. All images and graphs are representative of 3 mice per group. Quantification was performed across the entire tissue section and expressed as YFP+ cells per unit area. Data are mean ± s.d. Exact P-values are indicated. Illustration in a was created using BioRender ( https://biorender.com ).

Article Snippet: Slides were cover-slipped with EverBrite TrueBlack Hardset Mounting Medium with DAPI (Biotium, 23018) or ProLong Gold Antifade Mountant (Invitrogen, P36930 ) after nuclear staining with 1 μg ml −1 DAPI (Thermo Scientific, 62248) for 10 min at room temperature.

Techniques: Immunofluorescence, Control, In Vivo, Staining

Journal: Nature

Article Title: The transition from monocyte to tissue-resident macrophage requires DHPS

doi: 10.1038/s41586-025-09972-2

Figure Lengend Snippet: a , Heat map of stable transcripts reduced on ribosomes in peritoneal macrophages (pMacs) ( Dhps- ΔM). b – d , pMacs analysed for IL33R (ST2) ( b ); cultured with IL-33 (20 ng ml −1 ) for 72 h or IL-4 (20 ng ml −1 ) for 24 h and analysed ( c ); or analysed for IL-4R ( d ). e , Bulk RNA-seq scheme. f , Volcano plot of shared DEGs. g , DAVID pathway analysis of downregulated genes in Dhps -deficient macrophages. h , L1CAM and E-cadherin, flow cytometry; pMacs from Dhps +/+ Rosa26 eYFP and Dhps -ΔM Rosa26 eYFP mice. i , Percentage cell recovery by time after EDTA; pMacs were cultured for 24 h. j , k , Confocal images of cultured pMacs with cell structure by brightfield microscopy ( j ) (arrows indicate rounded cells) and lungs stained for macrophages and stromal cells (F4/80, CD64, and PDGFRa, respectively) from Dhps -WT and Dhps -ΔM mice ( k ). Nuclei (DAPI); box indicates region of interest. l , Area and sphericity of F4/80 + CD64 + macrophages from k . Each data point is the average of five random fields quantified per biological replicate. m , Lungs from Dhps -WT and Dhps -ΔM mice stained for macrophages (F4/80, brown), vasculature (CD31, purple), epithelial cells (EpCAM, teal) and nuclei (haematoxylin, indigo). Frequency of F4/80 + cells not positioned on tissue (non-overlapping) was quantified from five random fields in three biological replicates per genotype. n , Confocal images of YFP + kidney macrophages from Dhps +/+ (control) and Dhps flx/flx CX3CR1–ERT2cre-Rosa26 eYFP mice 5 weeks post-tamoxifen. Cell volume and sphericity were quantified by Imaris (10.0.1, Bitplane); five random fields in five biological replicates per genotype. All data represent two or more experiments, except in a and e – g , in which data represent one experiment with three biological replicates. Data are mean ± s.d. For b and c , n = 3, Dhps -WT; and n = 4, Dhps -ΔM. For d , n = 5, Dhps -WT; and n = 4, Dhps -ΔM. For h , i , l and m , n = 3. For n , n = 5. n represents biological replicates. Statistical analyses; two-tailed t-tests and two-way analyses of variance; P values are shown. Scale bars, 50 μm ( k , top), 20 μm ( k , bottom); 20 μm ( n ). Illustrations in a , b , e and h – n were created using BioRender ( https://biorender.com ).

Article Snippet: Slides were cover-slipped with EverBrite TrueBlack Hardset Mounting Medium with DAPI (Biotium, 23018) or ProLong Gold Antifade Mountant (Invitrogen, P36930 ) after nuclear staining with 1 μg ml −1 DAPI (Thermo Scientific, 62248) for 10 min at room temperature.

Techniques: Cell Culture, RNA Sequencing, Flow Cytometry, Cell Recovery, Microscopy, Staining, Control, Two Tailed Test

Journal: Frontiers in neurology

Article Title: Inside the Thrombus: Association of Hemostatic Parameters With Outcomes in Large Vessel Stroke Patients.

doi: 10.3389/fneur.2021.599498

Figure Lengend Snippet: FIGURE 3 | Matrix metalloproteinase-10 (MMP-10) and thrombin-activatable fibrinolysis inhibitor (TAFI) colocalize with inflammatory cells and platelets in thrombi. Double immunofluorescence for MMP-10 (top, red) and TAFI (bottom, red) and leukocytes (CD45, left), macrophages CD68 (middle), and platelets CD42b (right, green); cell nuclei are stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Arrow heads point to double positive cells for MMP-10 (upper panels) and TAFI (lower panels) and the specified antigens (yellow). Scale = 20 µm.

Article Snippet: Finally, slides were mounted with VECTASHIELD R© Antifade Mounting Medium on DAPI (Novus Biological).

Techniques: Staining

Journal: Frontiers in neurology

Article Title: Inside the Thrombus: Association of Hemostatic Parameters With Outcomes in Large Vessel Stroke Patients.

doi: 10.3389/fneur.2021.599498

Figure Lengend Snippet: FIGURE 4 | Matrix metalloproteinase-10 (MMP-10) and thrombin-activatable fibrinolysis inhibitor (TAFI) colocalization in thrombi. Immunofluorescence for TAFI (red), MMP-10 (green), and 4′,6-diamidino-2-phenylindole (DAPI) (blue). Arrow heads point to double positive cells for MMP-10 and TAFI. Scale = 20 and 10 µm.

Article Snippet: Finally, slides were mounted with VECTASHIELD R© Antifade Mounting Medium on DAPI (Novus Biological).

Techniques: