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. 2015 May 6;2:15013. doi: 10.1038/mtm.2015.13

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Copyright © 2015 American Society of Gene & Cell Therapy

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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Expression of proNGF is dependent on p75^NTR in EC. Human retinal ECs were transduced with one of the followings: pGFP plus scrambled siRNA (GFP+Scr), pGFP plus siRNA against p75^NTR (GFP+siRNA); pGFP-proNGF123, a mutant cleavage resistant form of proNGF, plus scrambled siRNA (proNGF+Scr); pGFP-proNGF123 plus siRNA against p75^NTR (proNGF+siRNA), and cell lysate and supernatant were collected after 48 hours. (a,b) Representative western blots statistical analysis of total p75^NTR expression normalized to actin. (a,c) Representative western blots and statistical analysis of proNGF expression normalized to actin. 2 × 2 analysis showed interaction between proNGF overexpression and silencing p75^NTR. Silencing p75^NTR significantly reduced its expression in both GFP-controls and proNGF-groups. Results showed that overexpression of proNGF induced p75^NTR expression and that silencing p75^NTR downregulated proNGF expression in EC. Results presented as mean ± SD. n = 5–6 per group. *P < 0.05 versus other groups. (d) Representative western blots and statistical analysis of NGF expression normalized to actin in EC. (e) Representative western blots and statistical analysis of TrkA activation normalized to total TrkA. 2 × 2 analysis demonstrated interaction between proNGF overexpression and silencing p75^NTR. Overexpression of proNGF resulted in significant decreases in NGF levels and activation of its TrkA receptor compared with GFP-controls. Silencing p75^NTR expression increased NGF levels and enhanced TrkA activation in proNGF overexpressing cells. Results are expressed as mean ± SD. n = 5 per group. *P < 0.05 versus other groups.