Figure 3
TRPC3 participates in SOCE in DRG neurons. (A) TRPC3 shRNA-mediated knockdown was validated in a functional assay to determine the inhibitory effect of the shRNA on OAG-evoked calcium entry in DRG neurons (n = 9-11, P < 0.0001). (B) shRNA-mediated knockdown of TRPC3 in the SOCE assay clearly shows a strong contribution of this channel to the overall calcium influx. A decrease of approximately 42% is observed (n = 13-27, P < 0.05). (C) Heterologous overexpression of TRPC3 (OE) in DRG neurons produced a drastic increase in SOCE-mediated calcium influx of approximately 85% (n = 16-19, P < 0.01). (D) The inhibition of TRPC3 activity by the selective blocker Pyr10 (10 μM, 15 min) also resulted in a strong decrease of SOCE activity, with calcium influx dropping approximately 50% (n = 34-39, P < 0.01). The cumulative inhibition of both Orai and TRPC3 with Gd3+ and Pyr10, respectively, almost completely abolished SOCE response in DRG (n = 18-21, P < 0.001).