prox1  (Novus Biologicals)

Journal: Computational and Structural Biotechnology Journal

Article Title: Rapid 3D Immunolabeling and Light Sheet Microscopy for Quantitative Analysis of Intact Tissues

doi: 10.34133/csbj.0121

Figure Lengend Snippet: Age-associated lymphatic vessel expansion in endocrine glands. (A) Whole-organ imaging of cleared mouse ovary, testis, and thyroid gland from young and aged mice immunostained with LYVE-1. Asterisks indicate higher magnification of specific regions. Quantification of lymphatic vessel density in young and aged ovary, testis, and thyroid gland ( n = 8). (B) 3D confocal images of young and aged ovary, testis, and thyroid gland with Podoplanin and Prox1 immunostaining. Nuclei: DAPI. Quantification of Prox1 + cell numbers in young and aged ovary, testis, and thyroid gland ( n = 8). (C) qPCR analysis of Prox1 expression (normalized to Actb ) in aged human ovary, testis, and thyroid compared with young endocrine glands ( n = 4). Data represent mean ± SD. P values were derived from 2-tailed unpaired t tests and corrected for multiple comparisons using the Benjamini–Hochberg false discovery rate procedure. **** P < 0.0001. Scale bars: 400 μm for whole-organ images and 50 μm for high-magnification insets (A) and 50 μm (B).

Article Snippet: Quantification of Prox1 + cell numbers was performed using Imaris software (version 9.2.1) based on single-cell resolution 3D images.

Techniques: Imaging, Immunostaining, Expressing, Derivative Assay

Journal: bioRxiv

Article Title: Somatic variants activating the RAS-MAPK pathway confer susceptibility to hippocampal sclerosis in drug-resistant epilepsy

doi: 10.64898/2026.04.06.716727

Figure Lengend Snippet: Variant allele fractions (VAFs) from amplicon sequencing were compared across brain regions and cell types (A) . In four of six cases with findings, the causal somatic variant was identified in both cortex and hippocampus tissue (B) . One case with a PTPN11 A461T variant exhibited extensive neuronal loss in the CA1 region, as seen in H&E and NeuN stains (C) . Fluorescence-activated nuclei sorting was used to isolate hippocampal cell types from this tissue for amplicon sequencing (D) . The PTPN11 A461T variant was present in all cell types but highly enriched among Prox1-neurons (E) .

Article Snippet: Nuclei were labeled using the following antibodies: NeuN (Millipore, #MAB377X), PROX1 (Novus Biologicals, #NBP1-30045AF647), OLIG2 (Novus Biologicals, # NBP2-89201AF594), and PAX6 (Novus Biologicals, #NBP2-34705AF647) at 1:100 followed by incubation for 1 hour at 4°C with agitation.

Techniques: Variant Assay, Amplification, Sequencing, Fluorescence

Journal: bioRxiv

Article Title: Somatic variants activating the RAS-MAPK pathway confer susceptibility to hippocampal sclerosis in drug-resistant epilepsy

doi: 10.64898/2026.04.06.716727

Figure Lengend Snippet: Sorting scheme to enrich astrocytes, oligodendrocytes, Prox1-negative neurons and Prox1-positive neurons for amplicon sequencing.

Article Snippet: Nuclei were labeled using the following antibodies: NeuN (Millipore, #MAB377X), PROX1 (Novus Biologicals, #NBP1-30045AF647), OLIG2 (Novus Biologicals, # NBP2-89201AF594), and PAX6 (Novus Biologicals, #NBP2-34705AF647) at 1:100 followed by incubation for 1 hour at 4°C with agitation.

Techniques: Amplification, Sequencing

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