Figure 2
Magnetic stimulation of EPG in HEK293T cells induced significant increases in [Ca2+]i. (A) HEK293T cells transfected with pLV-CMV-EPG-IRES-hrGFP. (B,C) In vitro calcium images of fura-2/AM loaded cells before (B) and after (C) 10 s of magnetic stimulation. Black arrow indicates cell body. Scale bar 10 µM. (D) Raw data showing the fluorescence increase after magnetic stimulation (blue bar) in cells transfected with EPG. (E) Raster plot of transfected cells responding to magnetic stimulation (red dots, n = 68). Cells were classified as responders if the fluorescence signal peak was 10 SD above the mean. (F) Data showing kinetics of fluorescence intensity response for individual cells under different conditions. Decreased fluorescence intensity changes were observed in 0 Ca2+ and 0 Ca2+ + EGTA after magnetic stimulation. No increase in signal was observed for control (non-transfected) and thapsigargin treated cells. (G) Data showing the average (±SEM) changes in fluorescence before and after magnetic stimulation for each condition n = 68 (EPG), n = 76 (Control), n = 23 (0 Ca2+), n = 9 (0 Ca2+ + EGTA) & n = 48 (Thapsigargin).One-way ANOVA, Dunnett’s post-test (F4, 219) = 27.47, P < 0.0001.
