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. 2011 May 31;108(24):9747–9752. doi: 10.1073/pnas.1102962108

Fig. 4.

Fig. 4.

CS-E modulates NGF- or NT-4/5-mediated TrkA activation in cells. (A) Removal of endogenous CS-E on PC12 cells using ChABC reduced NGF- and NT-4/5-mediated TrkA phosphorylation. (B) CS-E-enriched polysaccharides enhanced TrkA phosphorylation by NT-4/5, but not NGF, when coated on a substratum at 500 ng·mL-1. (C) Addition of exogenous CS-E (500 ng·mL-1) to the media reduced NGF-induced and NT-4/5-induced TrkA phosphorylation. NGF-induced activation was further inhibited by higher concentrations (5,000 ng·mL-1) of CS-E. For A–C, relative TrkA activation was calculated with respect to total TrkA levels for each condition and was plotted relative to that of untreated cells in the presence of the indicated NT. n = 4, *P < 0.05, **P < 0.005. (D) Prolonged NGF treatment (60 min), increases the colocalization of CS-E and TrkA. Representative images show minimal colocalization in untreated cells and increased colocalization (yellow) after treatment with NGF. The extent of colocalization was quantified as described in SI Methods and plotted relative to that of untreated cells. n = 24 cells. **P < 5 × 10-6.