ccl21a  (Vector Laboratories)

Journal: eLife

Article Title: Transcriptomic profiling of Schlemm’s canal cells reveals a lymphatic-biased identity and three major cell states

doi: 10.7554/eLife.96459

Figure Lengend Snippet: ( A ) scRNA-seq of 129/Sj anterior segment tissue identifies various cell types similar to that in C57BL/6 J single-cell RNA sequencing (left panel). Expression of Egfl7 and Cdh5 in snRNA-seq endothelial cells (right panel). ( B ) Integration of B6 and 129/Sj endothelial cells followed by sub-clustering identifies BECs, LECs, IW1 SECs, IW2 SECs, OW SECs, and CCs (top panel). Integration of B6 and 129/Sj endothelial cells distributed across clusters (bottom panel). ( C ) Sub-clustering identifies complementary expression patterns of Npnt and Ccl21a in IW1 and IW2 SECs, Ackr1 in CCs, and Selp in OW SECs. ( D ) Heatmap of differentially expressed genes of the identified sub-clusters. ( E ) Violin plot showing differences in expression levels of various genes which as a combination defines individual sub-clusters. IW: Inner wall, OW: Outer wall, CC: Collector channels, BEC: Blood endothelial cell, LEC: Lymphatic endothelial cell, SEC: Schlemm’s Canal endothelial cell.

Article Snippet: Antibody , CCL21A (Goat polyclonal) , R&D Systems , Cat# AF457-SP; RRID: AB_2072083 , WM (1:25) Sections (1:200).

Techniques: RNA Sequencing Assay, Expressing

Journal: eLife

Article Title: Transcriptomic profiling of Schlemm’s canal cells reveals a lymphatic-biased identity and three major cell states

doi: 10.7554/eLife.96459

Figure Lengend Snippet: ( A ) Npnt expression in a subgroup of SEC in scRNA-seq data (i) and corresponding immunofluorescence (IF) reveals high level of expression of NPNT in anterior portion of IW of SC in a frozen section (ii) and whole mount (iii and iv). ( B ) Ccl21a is expressed in SECs and LECs (i) and corresponding IF reveals high expression in posterior portion of IW of SC in a frozen section (ii) and whole mount (iii and iv). ( C ) Selp is expressed in OW SECs and CCs, a subgroup of SECs in single-cell (i) and corresponding IF (ii frozen section, iii-iv whole mount). ( D ) Ackr1 expression in a subset of CC cells (i) and corresponding IF (ii frozen section, iii whole mount). DAPI in blue labels nuclei in all panels. IW: Inner wall, OW: Outer wall, CC: Collector channels, CB: Ciliary body, LY: Lymphatic vessels, BV: Blood vessels SC: Schlemm’s canal. Ant.: Anterior SC, Post.: Posterior SC. Scale bar = 100μm.

Article Snippet: Antibody , CCL21A (Goat polyclonal) , R&D Systems , Cat# AF457-SP; RRID: AB_2072083 , WM (1:25) Sections (1:200).

Techniques: Expressing, Immunofluorescence

Journal: eLife

Article Title: Transcriptomic profiling of Schlemm’s canal cells reveals a lymphatic-biased identity and three major cell states

doi: 10.7554/eLife.96459

Figure Lengend Snippet: ( A ) Immunofluorescence (IF) of NPNT shows variability in expression with a gradient of high expression in the anterior portion of SC (left panel) and uniform expression throughout IW of SC (middle panel) in flash-frozen sections. In situ hybridization using RNAscope shows the expression of Npnt in IW with an anterior expression bias (left panel) ( B ) IF of CCL21A shows variability in expression with a gradient of high expression in the posterior portion of SC (right panel) and uniform expression throughout IW of SC (left panel) in flash-frozen sections. DAPI in blue labels nuclei in all panels. ( C, D ) Gene expression analysis of endothelial cell subset in published dataset shows similar segregation of Npnt, Selp, and Ccl21a expression in SECs as seen in our dataset. CB: ciliary body, SC: Schlemm’s canal. Ant.: Anterior Schlemm’s canal, Post.: Posterior Schlemm’s canal. Scale bar = 100μm.

Article Snippet: Antibody , CCL21A (Goat polyclonal) , R&D Systems , Cat# AF457-SP; RRID: AB_2072083 , WM (1:25) Sections (1:200).

Techniques: Immunofluorescence, Expressing, In Situ Hybridization, RNAscope

Journal: eLife

Article Title: Transcriptomic profiling of Schlemm’s canal cells reveals a lymphatic-biased identity and three major cell states

doi: 10.7554/eLife.96459

Figure Lengend Snippet:

Article Snippet: Antibody , CCL21A (Goat polyclonal) , R&D Systems , Cat# AF457-SP; RRID: AB_2072083 , WM (1:25) Sections (1:200).

Techniques: Transgenic Assay, Staining, Immunofluorescence, RNA Sequencing Assay

Journal: Arthritis Research & Therapy

Article Title: Lymphotoxin-beta receptor blockade reduces CXCL13 in lacrimal glands and improves corneal integrity in the NOD model of Sjögren's syndrome

doi: 10.1186/ar3507

Figure Lengend Snippet: Lacrimal gland gene expression reduced by treatment of mice with LTBR-Ig compared to control.

Article Snippet: Data obtained by multiplex real time PCR analysis, shown in Results, was performed using Taqman primer sets (CXCL9, CXCL12, CXCL13, CCL19, CCL20, CCL21a, Chst2, Chst4, GlyCAM-1, VCAM, MAdCAM) from ABI using the Applied Biosystems 7900HT PCR System (Carlsbad, CA, USA).

Techniques: Expressing, Sequencing

Journal: Arthritis Research & Therapy

Article Title: Lymphotoxin-beta receptor blockade reduces CXCL13 in lacrimal glands and improves corneal integrity in the NOD model of Sjögren's syndrome

doi: 10.1186/ar3507

Figure Lengend Snippet: CXCL13 protein measured by ELISA in mouse lacrimal glands and in sera from Sjögren's patients . Homogenates made from individual lacrimal glands ( n = 8) were analyzed for CXCL13 protein levels by ELISA (a,b) . CXCL13 protein increased with age/disease progression in lacrimal glands from untreated male NOD mice (a). LTBR-Ig treatment from 8 to 16 weeks reduced CXCL13 content of mouse lacrimal glands (b). Mean concentration of CXCL13 was significantly elevated in serum samples of Sjögren's syndrome patients ( n = 27) versus healthy control sera ( n = 30) (c) . The mean age of Sjögren's syndrome patients was 57.17 +/- 10.10 years, and for controls was 57.55 +/- 21.17 years. There was one man in each group. ELISA, enzyme linked immunosorbant assay; NOD, non-obese diabetic.

Article Snippet: Data obtained by multiplex real time PCR analysis, shown in Results, was performed using Taqman primer sets (CXCL9, CXCL12, CXCL13, CCL19, CCL20, CCL21a, Chst2, Chst4, GlyCAM-1, VCAM, MAdCAM) from ABI using the Applied Biosystems 7900HT PCR System (Carlsbad, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay