Fig. 6: The NMB-NMBR ligation activates T-type calcium channels by PKA activation.

a The intracellular Ca²⁺ concentration was analyzed in Schwann cells treated with NMB and PD168368 by measuring the fluorescence intensity of the calcium indicator Fluo-4/AM with a confocal microscope. b The statistics of peak calcium responses were analyzed in a (one-way ANOVA and Tukey’s multiple comparisons test). c The intracellular Ca²⁺ concentration was analyzed in Schwann cells treated with NMB, PD168368, Nifedipine, NP118809, Trimethadione and NiCL2 by measuring the fluorescence intensity of the calcium indicator Fluo-4/AM with a confocal microscope. d The statistics of peak calcium responses were analyzed in c (one-way ANOVA and Tukey’s multiple comparisons test). e The intracellular Ca²⁺ concentration was analyzed in Schwann cells treated with KT-5720 by measuring the fluorescence intensity of the calcium indicator Fluo-4/AM with a confocal microscope. f The statistics of peak calcium responses were analyzed in e (unpair t-test). g The intracellular cAMP concentration was analyzed in RSC96 cells treated with NMB and PD168368 by ELISA (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). h The intracellular cAMP concentration was analyzed in RSC96 cells cocultured with HeLa or Me180 with or without PD168368 (n = 5, one-way ANOVA and Tukey’s multiple comparisons test). i CREB phosphorylation were examined in RSC96 cells treated with a concentration gradient of NMB. Data are shown as photographs from one representative of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.