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: Hippocampus

Article Title: Neuronal activity-related transcription is blunted in immature compared to mature dentate granule cells

doi: 10.1002/hipo.23515

Figure Lengend Snippet: Reduced immediate early gene (IEG) protein expression in immature DGCs. (a) Experimental timeline. Mice received tamoxifen (TAM) gavage for 7 days to label DGCs and were exposed to a novel environment (NE) to stimulate activation 5 or 13 weeks (wk) later. Untreated littermates collected directly from the home cage (HC) were used as a control. (b) Sample of durable GFP labeling 13 weeks after TAM in the DG. Scale bar: 20 μm. Yellow arrows indicate a cell triple labeled for GFP, ARC, and FOS. Dashed yellow line indicates the boundaries of the granule cell layer (GCL). (c) NE exposure induces significantly more FOS and ARC expression than the home cage control condition (n’s: HC = 6, 5-wk NE = 7, 13-wk NE = 7). (d) Quantification of active 5-week and 13-week DGCs in coronal tissue sections shows greater IEG expression in more mature cells (n’s: 5-wk = 7, 13-wk = 7). (e) Quantification of active 5-week and 13-week DGCs via flow cytometry (FACS) also shows greater IEG expression in more mature cells (n’s: Arc 5-wk = 7, Arc 13-wk = 7, Fos 5-wk = 10, Fos 13-wk = 9). (f) Co-expression of DG marker proteins PROX1 and CTIP2 occurs in the majority of GFP+ nuclei assessed by FACS, but co-expression continues to increase between 5 and 13 weeks (n’s: 5-wk = 10, 13-wk = 9). (g) Sample FACS gating strategy for identifying active immature DGCs. DGCs were identified by co-labeling with marker proteins PROX1 and CTIP2. Cell age was identified by GFP. Cell activation was assessed with ARC or FOS. (h) Experimental timeline for seizure treatment. Mice received TAM gavage for 7 days to label DGCs and were given pentylenetetrazole (PTZ) 5 or 13 weeks later. PBS-injected littermates served as staining controls. Mice were perfused following a 90-min observation period after the injection. (i) Top: Sample of FOS labeling following either PBS (left) or PTZ (right). Scale bar: 50 μm. Bottom: Sample of cell classification in PTZ-treated mice. Turquoise arrows indicate a cell triple labeled for GFP, CTIP2, and FOS. Yellow arrows indicate a cell double labeled for GFP and FOS, negative for CTIP2. Dashed yellow line indicates GCL boundaries. Scale bar: 20 μm. (j) Quantification reveals that CTIP2−GFP+ immature cells activate at significantly lower rates than CTIP2+GFP+ immature cells. CTIP2+GFP+ immature cells still have reduced FOS relative to mature CTIP2+GFP− cells at 5 weeks after TAM even under seizure conditions (n = 4, within-subject comparison). Points and n’s in dot plots represent mice. Error bars are mean ± S.E.M. *p < .05, **p < .01, ***p < .001, ns = not significant.

Article Snippet: Mouse anti-PROX1 (EMD Millipore #MAB5654, RRID:AB_2170714).

Techniques: Expressing, Activation Assay, Labeling, Flow Cytometry, Marker, Injection, Staining, Comparison