goat anti mouse prox1  (R&D Systems)

Journal: bioRxiv

Article Title: A Csf1r lineage gives rise to dermal lymphatic endothelial cells

doi: 10.64898/2026.03.17.712362

Figure Lengend Snippet: A-C Bulk RNA-seq analysis of tdTomato (TOM)+ and TOM- endothelial cells (ECs) from whole E12.5 Csf1r-iCre;Rosa tdTom mouse embryos (dataset GSE117978). A Volcano plot of significantly differentially expressed transcripts with more than 100 counts per transcript; relevant genes are named; grey and red data points represent transcripts with at least twofold over- or under-representation, respectively ( i.e. , red data points are over-represented and grey data points under-represented in TOM+ ECs compared to TOM-ECs). B,C Relative expression levels for markers typical of lymphatic endothelial cells (LECs) ( B ) or tdTomato and Csf1r ( C ); mean ± SD, n = 3 embryos; NS, not significant, *P < 0.05, **P < 0.01 (Benjamini–Hochberg’s multiple comparisons test for P value adjustment, Padj). D RT-qPCR expression analysis for the indicated genes relative to Actb , showing fold change expression in TOM+ ECs relative to TOM- ECs; mean ± SD, n = 3 embryos; *P < 0.05, **P < 0.01 (two-tailed unpaired t-test). E Immunofluorescence staining with the indicated markers of E15.5 Csf1r-iCre ; Rosa tdTom dorsal dermis (scale bar: 200 μm); the square indicates an area shown at higher magnification in the adjacent panels (scale bar: 25 μm), and shown for the different markers also in grey scale. F Immunofluorescence staining with the indicated markers for quantification of TOM+ LECs in E13.5 and E15.5 Csf1r-iCre ; Rosa tdTom dermis (scale bars: 100 μm); the squares indicate areas shown at higher magnification in the adjacent panels. The bar plot shows the fraction of TOM+ PROX1+ cells in E13.5 and E15.5 Csf1r-iCre ; Rosa tdTom dermis. Mean ± SD, n = 4 embryos; each dot represents the value for one embryo. G Immunofluorescence staining with the indicated markers of adult Csf1r-iCre ; Rosa tdTom ear dermis (scale bar: 100 μm). The square indicates an area shown at higher magnification in the adjacent panel, and shown for the different markers also in grey scale. Arrows indicate TOM+ LECs; arrowheads indicate TOM+ macrophages; curved arrows indicate TOM+ BECs; empty symbols indicate lack of expression for the indicated marker.

Article Snippet: Embryonic dorsal dermis was dissected from formaldehyde-fixed embryos and then incubated in PBS containing 2% serum-free protein block (DAKO), 2% bovine serum albumin and 0.4% Triton X-100 before staining with a combination of the following primary antibodies: goat anti-mouse NRP2 (R&D Systems #AF567, 1:100), rabbit anti-mouse PROX1 (Biolegend # 925202, 1:50), goat anti-mouse PROX1 (R&D Systems # AF2727, 1:100), rabbit anti-mouse LYVE1 (Angiobio # 11-034, 1:100), rat anti-mouse PECAM1 (BD Pharmigen # 553370, 1:50), rat anti-mouse EMCN (Santacruz # sc-65495, 1:50), goat anti-mouse FLT4 (R&D Systems # AF743, 1:100), rat anti-mouse TER119 (Biolegend # 116241, 1:100), rabbit anti-mouse GFP (MBL # 598, 1:200), goat anti-mouse TOM (Origene #AB1140-100, 1:250), or rat anti-mouse TOM (Chromotek #5F8, 1:200).

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR, Two Tailed Test, Immunofluorescence, Staining, Marker

Journal: bioRxiv

Article Title: A Csf1r lineage gives rise to dermal lymphatic endothelial cells

doi: 10.64898/2026.03.17.712362

Figure Lengend Snippet: A Representative immunofluorescence staining with the indicated markers and quantification of TOM+ LECs of E15.5 Csf1r-iCre ; Rosa tdTom dermis (scale bar: 100 μm) from mice with ( Spi1 +/+ ) or without ( Spi1 -/- ) differentiated myeloid cells. The square indicates an area shown at higher magnification in the adjacent panel. The bar plot shows the fraction of TOM+ PROX1+ cells; mean ± SD, n = 5 Spi1 -/- , n = 3 Spi1 +/+ embryos; each dot represents the value from one embryo. B Immunofluorescence staining with the indicated markers of E17.5 Spi1 -/- ; Csf1r-iCre ; Rosa Yfp dermis (scale bar: 100 μm); the square indicates an area shown at higher magnification in the adjacent panel, and shown for the different markers also in grey scale. Arrows indicate TOM+ LECs; arrowheads indicate TOM+ macrophages; curved arrows indicate TOM+ BECs; empty symbols indicate lack of expression for the indicated marker.

Article Snippet: Embryonic dorsal dermis was dissected from formaldehyde-fixed embryos and then incubated in PBS containing 2% serum-free protein block (DAKO), 2% bovine serum albumin and 0.4% Triton X-100 before staining with a combination of the following primary antibodies: goat anti-mouse NRP2 (R&D Systems #AF567, 1:100), rabbit anti-mouse PROX1 (Biolegend # 925202, 1:50), goat anti-mouse PROX1 (R&D Systems # AF2727, 1:100), rabbit anti-mouse LYVE1 (Angiobio # 11-034, 1:100), rat anti-mouse PECAM1 (BD Pharmigen # 553370, 1:50), rat anti-mouse EMCN (Santacruz # sc-65495, 1:50), goat anti-mouse FLT4 (R&D Systems # AF743, 1:100), rat anti-mouse TER119 (Biolegend # 116241, 1:100), rabbit anti-mouse GFP (MBL # 598, 1:200), goat anti-mouse TOM (Origene #AB1140-100, 1:250), or rat anti-mouse TOM (Chromotek #5F8, 1:200).

Techniques: Immunofluorescence, Staining, Expressing, Marker

Journal: bioRxiv

Article Title: A Csf1r lineage gives rise to dermal lymphatic endothelial cells

doi: 10.64898/2026.03.17.712362

Figure Lengend Snippet: A,B Strategy for the combined Csf1r-iCre -mediated lineage tracing and targeting of Prox1 ( A ) and representative immunofluorescence staining with the indicated markers of E15.5 dermis from a heterozygously targeted Csf1r-iCre ; Prox1 fl(Egfp)/+ mouse ( B ) (scale bars: 25 μm). C,D Representative images of E15.5 Prox1 fl/fl embryos with or without Csf1r-iCre (scale bars: 1 mm) ( C ) and table showing the frequency of embryos displaying the indicated phenotype ( D ); n = 20 Prox1 fl/+ , n = 22 Prox1 fl/fl , n = 22 Csf1r-iCre ; Prox1 fl/+ , n = 19 Csf1r-iCre ; Prox1 fl/fl from 11 litters. E Immunofluorescence staining with the indicated markers of E15.5 dermis of Csf1r-iCre ; Rosa tdTom ; Prox1 fl(Egfp)/+ and Csf1r-iCre ; Rosa tdTom ; Prox1 fl(Egfp)/fl(Egfp) littermate mice identifies erythrocytes in mutant dermal lymphatic vessels that are co-labelled for TOM and GFP (scale bars: 100 μm). F Representative immunofluorescence staining with the indicated markers of E15.5 Prox1 fl(Egfp)/fl(Egfp) (no Cre, normal PROX1 function), Csf1r-iCre ; Prox1 fl(Egfp)/+ (heterozygous PROX1 deficiency) and E15.5 Csf1r-iCre ; Prox1 fl(Egfp)/fl(Egfp) (homozygous PROX1 deficiency) dermis illustrates that TER119+ erythrocytes are located in NRP2+ lymphatic vessels of PROX1-deficient embryos. The square indicates an area shown at higher magnification in the adjacent panels and shown for the different markers also in grey scale (Scale bars: 100 μm). Arrows indicate TER119+ erythrocytes in lymphatic vessels.

Article Snippet: Embryonic dorsal dermis was dissected from formaldehyde-fixed embryos and then incubated in PBS containing 2% serum-free protein block (DAKO), 2% bovine serum albumin and 0.4% Triton X-100 before staining with a combination of the following primary antibodies: goat anti-mouse NRP2 (R&D Systems #AF567, 1:100), rabbit anti-mouse PROX1 (Biolegend # 925202, 1:50), goat anti-mouse PROX1 (R&D Systems # AF2727, 1:100), rabbit anti-mouse LYVE1 (Angiobio # 11-034, 1:100), rat anti-mouse PECAM1 (BD Pharmigen # 553370, 1:50), rat anti-mouse EMCN (Santacruz # sc-65495, 1:50), goat anti-mouse FLT4 (R&D Systems # AF743, 1:100), rat anti-mouse TER119 (Biolegend # 116241, 1:100), rabbit anti-mouse GFP (MBL # 598, 1:200), goat anti-mouse TOM (Origene #AB1140-100, 1:250), or rat anti-mouse TOM (Chromotek #5F8, 1:200).

Techniques: Immunofluorescence, Staining, Mutagenesis

Journal: bioRxiv

Article Title: Mitotic bookmarking by Prox1 preserves mammalian neuronal lineage identity memory via promoting timely H3K27me3 restoration

doi: 10.64898/2026.02.25.707603

Figure Lengend Snippet: ( a ) Representative images of control ( Prox1 fl/fl ) and Prox1 Nes-cKO ( Nestin-CreER T2 ; Prox1 fl/fl ) hippocampus at P120 stained for Prox1 and NeuN (Neuronal nuclei), a pan-neuronal marker. Scale bar, 200 μm. ( b ) Representative swimming paths of the control and Prox1 Nes-cKO mice before and after training in Morris Water Maze test. ( c ) A schematic diagram showing CA and DG neuroepithelium near the ventricular surface, with the neurogenesis process (arrows) producing CA and DG neurons respectively. ( d–f ) Representative images of developing hippocampus at E13 stained for Prox1 and pH3 ( d , f ) and quantifications of the relative Prox1 expression levels from CA to DG ( e ; n =27 brain sections). High magnification of the boxed areas in ( d ) showing the distribution of Prox1 in dividing CA or DG NSCs ( f ). *p < 0.05, ****p < 0.0001; Student’s t-test. Scale bar, 50 μm ( d ); 25 μm ( f ). The cell body of CA and DG NSCs is encircled by dotted lines. ( g , h ) Sample images of CA and DG NSCs in interphase stained with anti-Prox1 antibody and DAPI ( g ) and quantifications of Prox1 foci per NSC ( h ). n = 47, 60 cells; ****p < 0.0001; Student’s t-test. Scale bar, 5 μm. In this and subsequent micrographs, the cell body of CA and DG NSCs is encircled by dashed lines. ( i–k ) Representative images of metaphase (top) and anaphase (bottom) CA (left) and DG (right) NSCs at E13 stained with anti-Prox1, anti-pH3 and DAPI ( i ) and quantifications of Prox1 fluorescent intensity (F.I.) on the chromosomes (chro.) or in the cytosol (cyto.) ( j , k ). Arrowheads indicate Prox1 associating with chromosomes. n = 7, 9, 11, 14 cells, respectively; NS, not significant, ****p < 0.0001, ***p < 0.001; Student’s t-test. Scale bar, 5 μm. ( l ) A schematic diagram showing Prox1 (orange) mitotic retention in DG but not CA NSCs and progenitors.

Article Snippet: Antibodies used in this study include goat anti-Prox1 (1:50, R&D Systems, AF2727), goat anti-Sox2 (1:200, R&D Systems, AF2018), rabbit anti-Prox1 (1:200, Abcam, Ab101851), rat anti-Tbr2 (1:200, Thermo Fisher Scientific, 14-4875-82), rabbit anti-NeuroD1 (1:200, Abcam, Ab213725), rabbit anti-NeuN (1:200, Abcam, Ab177487), rabbit anti-NeuroD2 (1:200, Abcam, Ab104430), rabbit anti-NeuroD6 (1:200, Abcam, Ab85824), rabbit anti-FOXG1 (1:200, Abcam, Ab18259), rat anti-Ctip2 (1:200, Abcam, Ab18465), rabbit anti-Calb2 (Calretinin) (1:200, Abcam, Ab244299) and rabbit anti-Calb1 (Calbindin) (1:200, Abcam, Ab108404).

Techniques: Control, Staining, Marker, Expressing

Journal: Nature Communications

Article Title: Breast cancer remodels lymphatics in sentinel lymph nodes

doi: 10.1038/s41467-025-64981-z

Figure Lengend Snippet: a Differential abundance testing using miloR. Neighborhoods are colored by their log fold abundance change between distant (blue) and metastatic (red) LNs. Non-differential abundance neighborhoods (FDR >10%) are colored white. b Beeswarm plot of the cell subset distribution of log fold change between normal and distant LNs. c The traje c tories of LN LEC differentiation are shown in a UMAP plot. Six distinct LEC trajectories (T1-T6) were identified using Monocle single-cell trajectory analysis. d Immunostaining of cancer cells and LEC markers PROX1 and MARCO in non-metastatic (left) and metastatic LNs (right). Zoomed-in images displaying SCS containing cytokeratin + cancer cells (left) and medullary sinuses (MS), both with and without cancer cells (right). A cancer cell in the MARCO + sinus is indicated by an arrow. Blue, cytokeratin; red, PROX1; green, MARCO. Images are representatives of two individuals with similar results. e Immunostaining of CD200 + lymphatics and its quantification in metastatic (upper and lower, black background) and non-metastatic (lower, gray background) LNs. CD200 + lymphatics and individual cancer cells within lymphatics are indicated by arrows and arrowheads, respectively. Blue, cytokeratin; green, PROX1; red, CD200. Scale bars, 500 μm. Images are representatives of seven individuals with similar results. SCS: subcapsular sinus; MS: medullary sinus; B: B cell zone. Circles in the bar plots represent biological replicates (mean ± SEM, two-tailed, unpaired Student’s t -test). Metastatic LN, n = 5; non-metastatic LN, n = 3. Source data are provided as a Source Data file. Scale bars: 500 μm (zoomed-out), 100 μm (zoomed-in).

Article Snippet: The following primary antibodies were used: goat anti-human PROX1 (R&D Systems AF2727, 2 μg/ml), rabbit anti-human MARCO (Atlas Antibodies, HPA063793, 3 μg/ml), AF488 mouse anti-pan-cytokeratin (Thermo Fisher Scientific, MA5-18156, 1 μg/ml), mouse anti-human MGP (Novus Biologicals, NBP2-45844, 10 μg/ml), and BV421 mouse anti-human CD200 (Biolegend, 329209, 2 μg/ml).

Techniques: Immunostaining, Two Tailed Test

Journal: Nature Communications

Article Title: Breast cancer remodels lymphatics in sentinel lymph nodes

doi: 10.1038/s41467-025-64981-z

Figure Lengend Snippet: a Cellular subsets in non-metastatic and metastatic LNs from two patients (patient #3, 5). UMAP plots were generated from the Visium HD dataset and key cellular subsets, including LECs, cancer cells and immune cell subsets, were identified and spatially visualized within the tissue sections. The capsules were outlined by dotted lines. Scale bars, 500 μm. b UMAP showing subclusters of LEC from nonmetastatic and metastatic LNs. PROX1 + LECs from all samples were selected, integrated, and subclustered. c Spatial distribution of the LEC subsets identified in UMAP in both nonmetastatic and metastatic LNs. d Gene signatures of LEC subsets found in spatial transcriptomics.

Article Snippet: The following primary antibodies were used: goat anti-human PROX1 (R&D Systems AF2727, 2 μg/ml), rabbit anti-human MARCO (Atlas Antibodies, HPA063793, 3 μg/ml), AF488 mouse anti-pan-cytokeratin (Thermo Fisher Scientific, MA5-18156, 1 μg/ml), mouse anti-human MGP (Novus Biologicals, NBP2-45844, 10 μg/ml), and BV421 mouse anti-human CD200 (Biolegend, 329209, 2 μg/ml).

Techniques: Generated, Capsules

Journal: Nature Communications

Article Title: Breast cancer remodels lymphatics in sentinel lymph nodes

doi: 10.1038/s41467-025-64981-z

Figure Lengend Snippet: a Differential abundance testing using MiloR and a heatmap of differentially expressed genes between differential abundance neighborhoods in LN LECs. The UMAP plot was shown in Fig. and is inserted here for clarity. In the heatmap, columns and rows represent neighborhoods and differentially expressed genes, respectively. Expression values for each gene are scaled between 0 and 1. The upper panel of the heatmap shows the neighborhood log fold change. FDR, false discovery rate. b Heatmap showing the expression of the top DEGs between metastatic and distant LNs for each cell. Bars above the heatmap indicate the tissue and cluster origin of each cell (LNs, clusters). c Violin plots displaying the top DEG expression between metastatic and distant LNs by cluster, with log-normalized expression value labeled. Nine patients’ samples were integrated for this analysis ( a – c ). d Immunostaining of MGP and its quantification in metastatic and distant LNs. Zoomed-in images show SCS and medullary sinuses containing cancer cells and MGP expression on PROX1 + LECs (arrows). Blue, cytokeratin; red, MGP; green, PROX1. Scale bars, 125 μm. Images are representative of four individuals with similar results. Circles in the bar plots represent biological replicates (mean ± SEM, two-tailed, unpaired Student’s t -test). Metastatic LN, n = 4 patients; non-metastatic LN, n = 4 patients. Source data are provided as a Source Data file. e MGP expression in LECs of non-metastatic and metastatic LNs detected using Visium HD. f Gene Ontology (GO) enrichment analysis of the top DEGs (Wilcoxon rank-sum test, metastatic vs. distant LNs) using the EnrichR package. FDR-corrected p values are shown.

Article Snippet: The following primary antibodies were used: goat anti-human PROX1 (R&D Systems AF2727, 2 μg/ml), rabbit anti-human MARCO (Atlas Antibodies, HPA063793, 3 μg/ml), AF488 mouse anti-pan-cytokeratin (Thermo Fisher Scientific, MA5-18156, 1 μg/ml), mouse anti-human MGP (Novus Biologicals, NBP2-45844, 10 μg/ml), and BV421 mouse anti-human CD200 (Biolegend, 329209, 2 μg/ml).

Techniques: Expressing, Labeling, Immunostaining, Two Tailed Test