FIG 3.

TRPV2 activity enhances NGF-induced neurite outgrowth in PC12 cells. (A) Intracellular Ca²⁺ levels were measured by Fluo4 fluorescence using time-lapse microscopy. F11 cells expressing the empty vector (n = 80), WT TRPV2 (n = 72), or DN TRPV2 (n = 61) were treated with 2-APB (100 μM) at the 45-s time point. Data represent the mean normalized Fluo4 intensities ± SEM. (B) Representative micrographs of PC12 cells treated with NGF (100 ng/ml for 72 h) after transfection with GFP (control) or 1D4-tagged WT TRPV2 or DN TRPV2. After NGF treatment, cells were fixed and stained with 1D4 antibody followed by Alexa Fluor 488-labeled secondary antibody. Bar, 100 μm. (C) PC12 cells were transfected with GFP, WT TRPV2, or DN TRPV2; grown in the absence or presence of NGF (100 ng/ml for 72 h); and then imaged for fluorescence as described above for panel B. Data represent means ± SEM from 3 independent experiments. n.s., not significant. (D) Neurite lengths for PC12 cells transfected with GFP (n = 141), WT TRPV2 (n = 122), or DN TRPV2 (n = 67) and treated with NGF (100 ng/ml; 72 h). Data represent means ± SEM from 3 independent experiments.