Human Prox1 Antibody Best Seller

Detection of Mouse Prox1 by Western Blot

RNA-seq identifies the targets of GATA2 in primary human LECs. (A) Principal component analysis (PCA) was performed on RNA-seq data from control shRNA- and shGATA2-infected primary HLECs. A high level of similarity was observed within the groups as indicated by their proximity to each other. (B) Hierarchical clustering shows that approximately 1000 genes were consistently downregulated and 600 genes were upregulated in shGATA2-treated HLECs. (C) GO revealed a list of genes that are likely relevant to the phenotypes observed in mice lacking GATA2. (D) GATA2 was knocked out from a second HLEC line using CRISPR/Cas9. Western blot revealed the lack of GATA2 in the knockout cells (HLEC deltaGATA2). In contrast, no obvious differences were observed in the expression of PROX1. Additionally, qRT-PCR revealed the downregulation of miR-126. (A) n=3 independent experiments per shRNA; (D) n=3 independent experiments (antibiotic selection, western blot and qRT-PCR). **P<0.01. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31582413), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Prox1 by Western Blot

Prox1 regulates myoblast differentiation.(a) Analysis of Prox1 mRNA and protein during C2C12 myoblast differentiation (indicated by the increase in MyoD1 mRNA). (b) Prox1 protein and mRNA in human myoblasts before and after differentiation. Primary myoblast lines from two different individuals were analysed. (c–f) Analysis of myoblast proliferation and differentiation-related CyclinD1, Myf5 and MyoD mRNAs in shProx1 and shScr transduced primary human myoblasts before (c) and after differentiation for 7d (d), as well as in mouse C2C12 myoblasts before (e) and after (f) differentiation. (g) Myosin staining of C2C12 cells after 7d of differentiation. Note that Prox1 silencing completely blocked myotube development, as only occasional cells expressed myosin and these cells were negative for shProx1-GFP. Data is presented as mean±s.e.m., n=3+3 biological replicates in a–g (all experiments were repeated three times). One-way analysis of variance with repeated measures followed by Tukey's posthoc test and Student's two-tailed unpaired t-test, *P<0.05, **P<0.01, ***P<0.001. Scale bars, 50 μm. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/ncomms13124), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Prox1 by Immunocytochemistry/Immunofluorescence

In situ localization of Ptx3-LECs and transition between Ptx3-LECs and Marco-LECs in human LNs. (A) Expression of CD36/Cd36 in LN LEC subsets of human and mouse. Dots indicate mean log-normalized transcript count. (B–D) Identification of CD36high Ptx3-LECs in human head and neck LNs by immunostaining. (B,C) Immunofluorescence of PROX-1, MARCO and CD36 (B), or PROX-1, LYVE-1 and CLEC4M (C). Zoomed-in images (inset marked by blue dotted lines) in (B) and (C) demonstrate CD36high LYVE-1+ paracortical sinuses (filled arrowhead). Scale bars = 500 μm (left panels) and 100 μm (right panel inset). (D) CD36high LYVE-1+ Ptx3-LECs (filled arrowhead) can be seen associated with MARCO+ CLEC4M+ Marco-LECs (empty arrowhead) in human LNs. Scale bars = 100 μm. CD36high Ptx3-LECs were detected in four out of seven human LNs. Images are representative of four biological replicates. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32426372), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Prox1 by Immunocytochemistry/Immunofluorescence

In situ localization of Ptx3-LECs and transition between Ptx3-LECs and Marco-LECs in human LNs. (A) Expression of CD36/Cd36 in LN LEC subsets of human and mouse. Dots indicate mean log-normalized transcript count. (B–D) Identification of CD36high Ptx3-LECs in human head and neck LNs by immunostaining. (B,C) Immunofluorescence of PROX-1, MARCO and CD36 (B), or PROX-1, LYVE-1 and CLEC4M (C). Zoomed-in images (inset marked by blue dotted lines) in (B) and (C) demonstrate CD36high LYVE-1+ paracortical sinuses (filled arrowhead). Scale bars = 500 μm (left panels) and 100 μm (right panel inset). (D) CD36high LYVE-1+ Ptx3-LECs (filled arrowhead) can be seen associated with MARCO+ CLEC4M+ Marco-LECs (empty arrowhead) in human LNs. Scale bars = 100 μm. CD36high Ptx3-LECs were detected in four out of seven human LNs. Images are representative of four biological replicates. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32426372), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Prox1 by Western Blot

Prox1 regulates the slow muscle fibre gene program in skeletal muscle.(a) MyHC I (Myh7) expression in muscle fibres (arrows) in AAV8-Prox1 and AAV8-Ctrl transduced TA muscle. (b) Expression of myosin heavy chain and calcium-signalling genes in AAV8-Prox1 and AAV8-Ctrl transduced TA muscle. Note that the expression of slow fibre -specific genes are upregulated and fast MyHC genes downregulated in Prox1 overexpressing muscles. (c) Heat map from GSEA analysis showing that Prox1 overexpression in TA muscle leads to expression of many soleus specific myofibrillar genes (red colour indicates positive and blue negative enrichment). (d) Effect of Prox1 deletion on soleus muscle gene expression (HSA-CreERT2;Prox1fl/fl mice). Note the increased expression of fast MyHCs Myh2 and Myh4 RNAs, whereas slow troponin isoform RNAs and Myh7 RNA are decreased. (e) Deletion of Prox1 in slow soleus muscle with AAV-Cre leads to marked downregulation of MyHC I protein. Data is presented as mean±s.e.m., n=5+5 in a,b, n=3+3 in c, n=4+4 in d,e. Student's two-tailed unpaired t-test, *P<0.05, **P<0.01, ***P<0.001. Both overexpression and deletion experiments were conducted two times on independent mouse cohorts. WT, wild-type. Scale bars, 50 μm. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/ncomms13124), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Prox1 by Immunocytochemistry/Immunofluorescence

In situ localization of Ptx3-LECs and transition between Ptx3-LECs and Marco-LECs in human LNs. (A) Expression of CD36/Cd36 in LN LEC subsets of human and mouse. Dots indicate mean log-normalized transcript count. (B–D) Identification of CD36high Ptx3-LECs in human head and neck LNs by immunostaining. (B,C) Immunofluorescence of PROX-1, MARCO and CD36 (B), or PROX-1, LYVE-1 and CLEC4M (C). Zoomed-in images (inset marked by blue dotted lines) in (B) and (C) demonstrate CD36high LYVE-1+ paracortical sinuses (filled arrowhead). Scale bars = 500 μm (left panels) and 100 μm (right panel inset). (D) CD36high LYVE-1+ Ptx3-LECs (filled arrowhead) can be seen associated with MARCO+ CLEC4M+ Marco-LECs (empty arrowhead) in human LNs. Scale bars = 100 μm. CD36high Ptx3-LECs were detected in four out of seven human LNs. Images are representative of four biological replicates. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32426372), licensed under a CC-BY license. Not internally tested by R&D Systems.