anti mouse slit3 antibodies  (Affinity Biosciences)

Journal: Investigative Ophthalmology & Visual Science

Article Title: AQP5 Regulates Corneal Nerve Distribution Through NETRIN-1 and SLIT3

doi: 10.1167/iovs.66.14.6

Figure Lengend Snippet: NETRIN-1 expression was significantly reduced, whereas SLIT3 expression was markedly elevated in the corneal epithelium of Aqp5 −/− mice compared to Aqp5 +/+ mice. ( A ) Heatmap illustrates differentially expressed neurotrophic and axon guidance factors in the corneal epithelium of Aqp5 +/+ and Aqp5 −/− mice. ( B ) Volcano plot analysis of differentially expressed neurotrophic and axon guidance factors between the corneal epithelium of Aqp5 +/+ and Aqp5 −/− mice, with upregulated genes highlighted in red and downregulated genes in blue . ( C ) Quantitative RT-PCR analysis of Ntn1 and Slit3 expression levels in the corneal epithelium of Aqp5 +/+ and Aqp5 −/− mice ( n = 3). ( D ) Representative immunofluorescence images of corneas stained with NETRIN-1 and SLIT3 ( green ) and DAPI ( blue ). Scale bars : 20 µm. ( E ) Regional quantification of positive immunofluorescence staining from images in ( D ) ( n = 3). ( F ) Western blot analysis of NETRIN-1, SLIT3, and ROBO4 protein expression in the corneal epithelium of Aqp5 +/+ and Aqp5 −/− mice. ( G ) Statistical evaluation of Western blot data ( n = 6). ( H ) Immunofluorescence staining of primary corneal epithelial cells isolated from Aqp5 +/+ and Aqp5 −/− mice, labeled with DAPI ( blue ), NETRIN-1, and SLIT3 ( green ). Scale bars : 25 µm. ( I ) Regional quantification of positive immunofluorescence staining from images in ( H ) ( n = 6). ( J ) The protein concentration of NETRIN-1 and SLIT3 in primary corneal epithelial cell supernatant was detected through ELISA ( n = 3). Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant NETRIN-1 (4 μg/mL; #1109-N1; R&D Systems, Minneapolis, MN, USA), SLIT3 (4 μg/mL; #9296-SL; R&D Systems), corneal epithelial extracts, or neutralizing antibodies against NETRIN-1 (4 μg/mL; #AF1109; R&D Systems) and SLIT3 (4 μg/mL; #AF3629; R&D Systems) were added for 48 hours to assess their effects on axon growth.

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Western Blot, Isolation, Labeling, Protein Concentration, Enzyme-linked Immunosorbent Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: AQP5 Regulates Corneal Nerve Distribution Through NETRIN-1 and SLIT3

doi: 10.1167/iovs.66.14.6

Figure Lengend Snippet: NETRIN-1 and SLIT3 affected neuronal axon growth. ( A ) Axonal growth status of Aqp5 +/+ and Aqp5 −/− TG neurons following treatment with recombinant mouse NETRIN-1 (rmNETRIN-1) and recombinant mouse SLIT3 (rmSLIT3). Scale bars : 50 µm. ( B ) Statistical analysis of the maximum branch lengths of TG neurons ( n = 6). Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant NETRIN-1 (4 μg/mL; #1109-N1; R&D Systems, Minneapolis, MN, USA), SLIT3 (4 μg/mL; #9296-SL; R&D Systems), corneal epithelial extracts, or neutralizing antibodies against NETRIN-1 (4 μg/mL; #AF1109; R&D Systems) and SLIT3 (4 μg/mL; #AF3629; R&D Systems) were added for 48 hours to assess their effects on axon growth.

Techniques: Recombinant

Journal: Investigative Ophthalmology & Visual Science

Article Title: AQP5 Regulates Corneal Nerve Distribution Through NETRIN-1 and SLIT3

doi: 10.1167/iovs.66.14.6

Figure Lengend Snippet: Effects of corneal epithelial TEs and NAbs on axon growth in TG neurons. ( A ) Left : Axon growth was assessed following the addition of Aqp5 −/− and Aqp5 +/+ corneal epithelial TEs to Aqp5 +/+ TG neurons. Scale bars : 50 µm. Right : Statistical analysis of the maximum branch length of axons shown in ( A , i ) ( n = 6). ( B ) Left : Axon growth was assessed following the addition of Aqp5 −/− and Aqp5 +/+ corneal epithelial TEs to Aqp5 −/− TG neurons. Scale bars : 50 µm. Right : Statistical analysis of the maximum branch length of axons shown in ( B , i ) ( n = 6). ( C ) Left : Axon growth following the simultaneous addition of Aqp5 −/− TEs or SLIT3 NAbs. Scale bars : 50 µm. Right : Statistical analysis of the maximum branch length of axons presented in ( C , i ) ( n = 6). ( D ) Left : Axon growth following the simultaneous addition of Aqp5 +/+ TEs or NETRIN-1 NAbs. Scale bars : 50 µm. Right : Statistical analysis of the maximum branch length of axons shown in ( D , i ) ( n = 6). Results of statistical analyses are expressed as mean ± SD. *** P < 0.001.

Article Snippet: Recombinant NETRIN-1 (4 μg/mL; #1109-N1; R&D Systems, Minneapolis, MN, USA), SLIT3 (4 μg/mL; #9296-SL; R&D Systems), corneal epithelial extracts, or neutralizing antibodies against NETRIN-1 (4 μg/mL; #AF1109; R&D Systems) and SLIT3 (4 μg/mL; #AF3629; R&D Systems) were added for 48 hours to assess their effects on axon growth.

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: AQP5 Regulates Corneal Nerve Distribution Through NETRIN-1 and SLIT3

doi: 10.1167/iovs.66.14.6

Figure Lengend Snippet: The transcription factor JUN-D bound to and regulated the expression of NETRIN-1 and SLIT3 in corneal epithelial cells. ( A ) Heatmap analysis of Ntn1 and Slit3 cotranscription factor expression levels in the corneal epithelium of Aqp5 +/+ and Aqp5 −/− mice. ( B ) Western blot analysis of JUN-D protein expression levels in the corneal epithelium of Aqp5 +/+ and Aqp5 −/− mice. ( C ) Quantitative statistical analysis of Western blot data ( n = 3). ( D ) ChIP-PCR analysis revealed significant enrichment of JUN-D in the promoter regions of Ntn1 and Slit3 . ( E ) Quantitative RT-PCR analysis was conducted to evaluate the expression levels of Ntn1 and Slit3 in corneal epithelial cells isolated from Aqp5 +/+ mice following 24-hour treatment with SR11302 ( n = 3). ( F ) Western blot analysis of NETRIN-1 and SLIT3 protein expression in the corneal epithelial cells of Aqp5 +/+ mice treated with SR11302. ( G ) Quantitative statistical evaluation of Western blot data ( n = 6). Statistical results are presented as mean ± standard deviation (SD). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant NETRIN-1 (4 μg/mL; #1109-N1; R&D Systems, Minneapolis, MN, USA), SLIT3 (4 μg/mL; #9296-SL; R&D Systems), corneal epithelial extracts, or neutralizing antibodies against NETRIN-1 (4 μg/mL; #AF1109; R&D Systems) and SLIT3 (4 μg/mL; #AF3629; R&D Systems) were added for 48 hours to assess their effects on axon growth.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Isolation, Standard Deviation