Fig. 3
The TAAR1-mediated IA suppression requires novel PKC isozymes. a and b The time course curve of IA changes induced by tyramine (0.1 µM) in TG neurons pretreated with cholera toxin (CTX, 0.5 µg/mL for 16 h, n = 9 cells, a) or pertussis toxin (PTX, 0.2 µg/ml for 16 h, n = 10 cells, b). Insets indicate the representative traces. The letters indicate the points used for sample traces. c Summary data showing the effect of tyramine on IA in TG neurons pre-incubated with CTX or PTX. **p < 0.01 vs. control, paired t test. d Protein expression of Go or Gi in TG cells treated with NC-siRNA or Go siRNA (Go-siRNA). The immunoblots are representative of the results of at least three independent experiments. **p < 0.01 vs. control, unpaired t test. e Summary data demonstrating that Go-siRNA treatment abrogated the 0.1 µM tyramine-induced IA response (n = 12 cells). *p < 0.05 vs. control + NC-siRNA, unpaired t test. f Protein expression of Gi or Go in TG cells treated with NC-siRNA or Gi-siRNA. The immunoblots are representative of the results of at least three independent experiments. ***p < 0.001 vs. NC-siRNA, unpaired t test. g Summary of results demonstrating that Gi-siRNA treatment did not affect the ability of tyramine (0.1 µM) to decrease IA (n = 11 cells). *p < 0.05 vs. control + NC-siRNA, #p < 0.05 vs. control + Gi-siRNA, unpaired t test. h Summary data demonstrating the effect of 0.1 µM tyramine on IA in cells dialyzed with 10 µM QEHA (n = 8 cells), 10 µM SKEE (n = 11 cells) or 10 µM gallein (n = 9 cells). i Summary data demonstrating the effect of 0.1 µM tyramine on IA in TG neurons pre-incubated with KT5720 (1 µM, n = 10 cells), chelerythrine chloride (1 µM, n = 11 cells), sotrastaurin (100 nM, n = 10 cells) or Go6976 (200 nM, n = 11 cells). Application of 1 µM KT5720 (n = 6 cells), 1 µM chelerythrine chloride (n = 6 cells), 100 nM sotrastaurin (n = 5 cells), or 200 nM Go6976 (n = 7 cells) alone did not affect IA. **p < 0.01 vs. control, paired t test. j and k The time course curve of IA changes induced by tyramine (0.1 µM) in small TG neurons dialyzed with BAPTA (20 µM, n = 11 cells, j) or EI-150 (100 µM, n = 10 cells, k). Insets indicate the representative traces. The letters indicate the points used for sample traces. l Summary data demonstrating the effect of 0.1 µM tyramine on IA indicated in panels j and k. **p < 0.01 vs. control, paired t test