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: Cancer Research

Article Title: A Multipotent PROX1 + Tumor Stem/Progenitor Cell Population Emerges during Intestinal Tumorigenesis and Mediates Radioresistance in Colorectal Cancer

doi: 10.1158/0008-5472.CAN-23-1851

Figure Lengend Snippet: Prox1 expression in epithelial cell clusters from WT intestine and Apc Min/+ tumors. A–I, Small intestines from WT mice and tumors from the small intestine of the Apc Min/+ mice were dissociated into single cells and subjected to scRNA-seq. Apc Min/+ scRNA-seq data were pooled from 127 tumors pooled from 8 individual experiments. Female and male mice aged 16 to 20 weeks old were used. The WT data were pooled from three mice. A, Aggregated Uniform Manifold Approximation and Projection (UMAP) of the Epcam + cell clusters. B and C, UMAPs of the WT ( B ) and Apc Min/+ ( C ) samples. D, Composite UMAP of WT (turquoise) and Apc Min/+ (red) samples. The tumor-specific clusters are encircled. E, Proportion of each cell type across the WT and Apc Min/+ tumors. Statistically significant differences in cell cluster compositions as tested with scCODA are denoted by * (FDR <0.05). F and G, Violin plot ( F ) and feature plot ( G ) showing Prox1 expression. H and I, Violin plot ( H ) and feature plot ( I ) showing the expression of Lgr5 . Turquoise arrowheads in F and H point to the WT clusters in which Prox1 ( F ) and Lgr5 ( H ) are highly expressed. Stem/prog, stem/progenitor.

Article Snippet: Goat anti–human PROX1 antibodies (R&D Systems, #AF2727, RRID: AB_2170716) were used for immunoprecipitation and subsequent mass spectrometry analysis.

Techniques: Expressing

Journal: Cancer Research

Article Title: A Multipotent PROX1 + Tumor Stem/Progenitor Cell Population Emerges during Intestinal Tumorigenesis and Mediates Radioresistance in Colorectal Cancer

doi: 10.1158/0008-5472.CAN-23-1851

Figure Lengend Snippet: Analysis of Prox1 + cells in the Apc Min/+ cell clusters. A–I, Tumors from small intestines of 16 to 20 weeks old female and male Apc Min/+ mice were dissociated into single cells and subjected to scRNA-seq. Apc Min/+ scRNA-seq experiments were repeated eight times, and the analysis consists of 127 tumors isolated from 20 mice. A, Uniform Manifold Approximation and Projection (UMAP) showing the Epcam + cell clusters. B, Feature plots showing the expression of Prox1 , Lgr5 , Mki67 , and Olfm4 . C, Quantification of Prox1 + cells and Lgr5 + cells in the indicated cell clusters. D, Velocities derived from scvelo (dynamical model) and CellRank RNA velocity kernel projected into an UMAP embedding. E, Macrostates of cellular dynamics computed by CellRank indicating initial (stem/prog 1, stem/prog 2, and stem/prog 4) and terminal (goblet 2, enterocytes 2, and enterocytes 9) cell clusters. F–H, Random walk–based visualizations computed by CellRank starting in stem/prog 1 ( F ), stem/prog 2 ( G ), and stem/prog 4 ( H ) clusters, showing the evolution of cellular states toward their terminal points. The dots indicate the initial (black) and terminal (yellow) points of the walk. I and J, Monocle3 analysis of the scRNA-seq data showing the trajectory ( I ) and the pseudotime ( J ) results. *, P < 0.05; **, P < 0.005; ***, P < 0.0005. Ent, enterocytes; stem/prog, stem/progenitor.

Article Snippet: Goat anti–human PROX1 antibodies (R&D Systems, #AF2727, RRID: AB_2170716) were used for immunoprecipitation and subsequent mass spectrometry analysis.

Techniques: Isolation, Expressing, Derivative Assay

Journal: Cancer Research

Article Title: A Multipotent PROX1 + Tumor Stem/Progenitor Cell Population Emerges during Intestinal Tumorigenesis and Mediates Radioresistance in Colorectal Cancer

doi: 10.1158/0008-5472.CAN-23-1851

Figure Lengend Snippet: Prox1 + tumor cells function as multipotent tumor cells. A–H, Prox1 or Lgr5 lineage tracing was induced in Apc Min/+ mice by a single tamoxifen (TAM) dose (2 or 4 mg). The mice were euthanized 3 or 7 days later, and their tumors from the small intestine were subjected to scRNA-seq analysis and immunofluorescence staining. The Prox1-LT day 3 scRNA-seq data are pooled from two individual experiments, which included altogether four mice, Prox1-LT day 7 data from two experiments and seven mice, and Lgr5-LT data from two experiments and four mice. A, Schematic of the experimental setup. B, Uniform Manifold Approximation and Projection of the aggregated Epcam + cell clusters. Prolif. stem/prog, proliferating stem/progenitor clusters. C, A violin plot showing tdTomato expression in the indicated clusters in the Lgr5-LT (blue), Prox1-LT D3 (orange), and in Prox1-LT D7 (red) samples. Prolif. 2, proliferating stem/progenitor 2 cluster. D–F, Feature plots showing the expression of Prox1 ( D ), Lgr5 ( E ), and tdTomato ( F ) in the day 7 samples. The percentages in F indicate the fraction of tdTomato + cells among the enterocytes (top right corner) and stem/progenitor 1 cells (bottom left corner). G and H, Tissue sections from the Prox1-LT ( G ) and Lgr5-LT ( H ) mice stained for DAPI (DNA), RFP (TDTOMATO, red), and LYZ1, DCLK1, CHRA, MUC2, or ALPI (all green). Examples of double-positive cells are encircled in yellow. Scale bar, 50 µm.

Article Snippet: Goat anti–human PROX1 antibodies (R&D Systems, #AF2727, RRID: AB_2170716) were used for immunoprecipitation and subsequent mass spectrometry analysis.

Techniques: Immunofluorescence, Staining, Expressing

Journal: Cancer Research

Article Title: A Multipotent PROX1 + Tumor Stem/Progenitor Cell Population Emerges during Intestinal Tumorigenesis and Mediates Radioresistance in Colorectal Cancer

doi: 10.1158/0008-5472.CAN-23-1851

Figure Lengend Snippet: PROX1 + cells are selected upon irradiation. A–C, Uniform Manifold Approximation and Projection (UMAP; A ) of colorectal cancer organoids from patient 1, with (IR) and without (Ctrl) 2 Gy irradiation, analyzed by scRNA-seq 1 day after irradiation. B, PROX1 expression shown in a feature plot. PROX1 + cells are indicated in red; their percentages are also indicated; color intensity marks the PROX1 transcript expression level. C, A violin plot showing the PROX1 expression level in the organoids. D, Organoids isolated from tumors of colorectal cancer patients 2, 3, and 4 were irradiated with a 5-Gy daily dose on 5 consecutive days. RNA was isolated 1 day after the last irradiation. The columns show the PROX1 versus HPRT1 RNA ratio, quantified by qPCR (Ctrl = 1). Each dot represents a biological replicate, consisting of three technical replicates. E and F, A similar irradiation protocol as in D was used for SW1222 colorectal cancer cells. E, PROX1 RNA 1 day after the last irradiation dose. Each dot represents one biological replicate, which consists of two to three technical replicates. F, Proportion of PROX1 + SW1222 cells 1 day after each consecutive irradiation dose determined from a cell aliquot stained for PROX1. Seven to eight biological replicates per sample and time point were analyzed. G–J, LS174T colorectal cancer cells of volume 1 × 10 6 were injected subcutaneously into NOD/SCID gamma mice (NSG) mice (two tumors per mouse) and irradiated using either of the two irradiation protocols indicated ( G and H ). I, Quantification of the percentage of PROX1 + cells in control and irradiated tumors. Each dot indicates a single tumor. The data were pooled from two independent experiments. J, Representative images of LS174T xenografts stained for PROX1 (red) and DNA (DAPI). Scale bar, 50 µm. *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.0001; ns, nonsignificant.

Article Snippet: Goat anti–human PROX1 antibodies (R&D Systems, #AF2727, RRID: AB_2170716) were used for immunoprecipitation and subsequent mass spectrometry analysis.

Techniques: Irradiation, Expressing, Isolation, Staining, Injection, Control

Journal: Cancer Research

Article Title: A Multipotent PROX1 + Tumor Stem/Progenitor Cell Population Emerges during Intestinal Tumorigenesis and Mediates Radioresistance in Colorectal Cancer

doi: 10.1158/0008-5472.CAN-23-1851

Figure Lengend Snippet: Inhibition of LIG4 decreases the survival of PROX1 + cells following irradiation. A–D, Organoids isolated from Apc Min/+ mice or from the constitutively deleted VApc Min/+ P∆/∆ mice were plated and irradiated with a single dose of 2 to 10 Gy. A, Schematic of the experimental setup. B, Quantification of the proportion of dead (nontransparent) organoids in nonirradiated and irradiated samples. The experiment was repeated three times, each with 3 to 4 technical replicates. C and D, Representative brightfield and green fluorescent (caspase-3/7) images of the indicated treatment groups. Red and blue circles surround some of the organoids that were considered dead and alive, respectively ( D ). Scale bar, 1,000 μm. E and F, Percentage of cells expressing the indicated DDR-related transcripts in scRNA-seq data from Prox1 -positive ( Prox1 transcript >0.1; red) and Prox1 -negative (Prox1 = 0; gray) cells in Apc Min/+ tumors ( E ) and human colorectal cancer (hCRC; F ) samples. Number of Prox1 + / Prox1 − cells: Apc Min/+ , 2691/10069; human colorectal cancer, 441/5496. In the Apc Min/+ data, there are eight dots, each representing distinct scRNA-seq experiments (details in legend). The human colorectal cancer data have been pooled from 15 patients (individual patient-specific dots are not shown, GSE200997 ). G, qPCR of ID1 and ID3 in shSCR transduced (gray) and in PROX1 -silenced LS174T cells (light red), and in HCT116 cells transfected with control (black) or PROX1 -overexpressing (red) lentiviruses. The experiment was repeated three times, each with three technical replicates; two PROX1 expression vectors were used to decrease the likelihood of off-target effects. H, The percentage of PROX1 + cells in LS174T cultures treated with 10 µmol/L of SCR7 pyrazine or DMSO starting 1 day before 2 Gy irradiation and analyzed 4 day after irradiation. Growth medium supplemented with SCR7 pyrazine was replenished every 3 days. Data were pooled from three individual experiments. Each dot represents a biological replicate. I, Representative images of PROX1-stained LS174T sections from xenografts of NOD/SCID gamma mice (NSG) mice treated with SCR7 pyrazine (20 mg/kg) or PBS every day, starting 1 day before 2 Gy irradiation. The mice were euthanized 2 days after irradiation. Scale bar, 50 μm. The average proportion ±SEM of PROX1 + cells in each treatment group is shown in the figures. Statistics are as follows: **, Ctrl + IR vs. Ctrl no IR, and ^, SCR7 pyrazine no IR vs. Ctrl IR and SCR7 pyrazine + IR vs. Ctrl IR; P < 0.05. Number of mice: Control no IR, 7; SCR7 pyrazine no IR, 7; Ctrl + IR, 7; and SCR7 pyrazine + IR, 8. *, P < 0.05; **, P < 0.005; ***, P < 0.0005.

Article Snippet: Goat anti–human PROX1 antibodies (R&D Systems, #AF2727, RRID: AB_2170716) were used for immunoprecipitation and subsequent mass spectrometry analysis.

Techniques: Inhibition, Irradiation, Isolation, Expressing, Transfection, Control, Staining

Journal: Cancer Research

Article Title: A Multipotent PROX1 + Tumor Stem/Progenitor Cell Population Emerges during Intestinal Tumorigenesis and Mediates Radioresistance in Colorectal Cancer

doi: 10.1158/0008-5472.CAN-23-1851

Figure Lengend Snippet: Multipotent Prox1 + tumor stem/progenitor cells are resistant to irradiation. A–G, Uniform Manifold Approximation and Projections (UMAP; A ), feature plots ( B ), transcript quantifications ( C and D ), violin plots ( E and F ), and Monocle3 analysis ( G ) based on scRNA-seq tumor data from control and irradiated Apc Min/+ mice. Apc Min/+ scRNA-seq experiments were repeated two to eight times per treatment group, each consisting of 18 to 127 tumors (control: 127; IR 1 day: 52, IR 2 days: 18, and IR 7 days: 65 tumors) from 3 to 20 mice (control: 20; IR 1 day: 9, IR 2 days: 3, and IR 7 days: 8 mice). Female and male mice aged 16 to 20 weeks old were used. A, UMAP of the Epcam + clusters in the indicated treatment groups. B, Feature plots indicating, in red, Prox1 + , Lgr5 + , Cdkn1a + , and Cdkn1c + cells in each treatment group. Color intensity marks the transcript expression level. C and D, Average percentage of Lgr5 + ( C ) and Prox1 + ( D ) cells ± SEM in the stem/progenitor 1, 2, and 4 clusters. E and F, Violin plots showing the expression level of Lgr5 ( E ) and Prox1 ( F ) in stem/progenitor 1 and 2 clusters in the indicated treatment groups. G, Trajectory and pseudotime Monocle3 analysis of the irradiated samples. H–J, Prox1 -lineage tracing was induced in Apc Min/+ mice by a single tamoxifen dose (2 or 4 mg). On the following day, the mice were either sham-irradiated or irradiated with 2 Gy. Two or six days after irradiation, the mice were euthanized and their tumors from the small intestine were subjected to scRNA-seq analysis and immunofluorescence staining. Both Prox1-LT IR day 3 and day 7 scRNA-seq data were pooled from three mice. H, Schematic of experimental setup. I, UMAPs of Epcam + cells in the indicated samples and feature plots showing the expression of tdTomato in the indicated samples in Epcam + cells. J, Feature plots presenting tdTomato expression in Prox1 -negative cells. Prolif. stem/prog, proliferating stem/progenitor cluster. *, P < 0.05; **, P < 0.005; ****, P < 0.0001.

Article Snippet: Goat anti–human PROX1 antibodies (R&D Systems, #AF2727, RRID: AB_2170716) were used for immunoprecipitation and subsequent mass spectrometry analysis.

Techniques: Irradiation, Control, Expressing, Immunofluorescence, Staining