RabGEF1/Rabex-5 Regulates TrkA-Mediated Neurite Outgrowth and NMDA-Induced Signaling Activation in NGF-Differentiated PC12 Cells
Fig 4
Reduced RabGEF1 expression decreases NGF-induced TrkA phosphorylation but not TrkA internalization.
A) CMV and AS stable transfectants were stimulated by NGF (50 ng/ml) for 30 min, 1 hour, 3 hours, or 5 hours. Total cell lysates were analyzed by Western blot using anti-phospho-TrkA and anti-TrkA antibody. B) CMV and AS stable transfectants were incubated with biotinylated anti-rat TrkA IgG antibody for the indicated times. Surface TrkA was assessed by streptavidin (SA)-APC fluorescence and analyzed by flow cytometry. Percentage TrkA internalization was calculated by subtracting mean fluorescence intensity at 2.5 min, 15 min, 30 min, or 60 min from mean fluorescence intensity at 0 min and dividing this number by mean fluorescence intensity at 0 min (x 100%). Data shown were pooled from four separate experiments.