Fig. 1. Activation of Toll-like receptor 2 (TLR2) in trigeminal ganglion (TG) neurons. (A, B) Representative traces (different colors indicate three individual neurons, A) and quantitative data of reproducible [Ca2+]i responses induced by Pam3CSK4 (1 µg/ml, 100 s/application) in TG neurons (n=7 neurons from n=4 mice; one-way ANOVA, p=0.5346, ns, B). (C, D) A representative trace (C) and quantitative data showing insignificant effects on the Pam3CSK4-induced [Ca2+]i responses after a 10 min pretreatment with 1 µM thapsigargin (Thap) (n=54 neurons from n=4 mice; one-way ANOVA with Bonferroni post hoc test, ****p<0.0001; 1st vs. 2nd+Thap: ****p<0.0001; 2nd+Thap vs. 3rd: p=0.9838, ns; 1st vs. 3rd: **p=0.0033, D). (E, F) A representative trace (E) and quantitative data showing significant abolished Pam3CSK4-induced [Ca2+]i responses after a 10 min pretreatment with a 0 Ca2+ bath solution (n=26 neurons from n=6 mice; one-way ANOVA with Bonferroni post hoc test, **p=0.0011; 1st vs. 2nd+0 Ca2+: ***p=0.0001; 2nd+0 Ca2+ vs. 3rd: ***p=0.0002; 1st vs. 3rd: p>0.9999, ns, F). (G, H) Representative traces showing Pam3CSK4 (1 µg/ml, 100 s)-induced [Ca2+]i responses in TRPV1-positive (a blue trace) and TRPV1-negative (a black trace) TG neurons (G) and quantitative data showing the percentage of CAP-positive (82.6%, n=19 of 24) and CAP-negative neurons (17.4%, n=4 of 24) in Pam3CSK4-positive neurons (H). n=2 mice. (I) The percentage of Pam3CSK4-responding neurons in the wild-type (WT) mice (approximately 5.7%, n=7 of 122 neurons, used the same data with panel B), TRPV1 KO mice (approximately 1.2%, n=3 of 232 neurons from n=2 mice), and TLR2 KO mice (0%, n=0 of 34 neurons from n=1 mouse). One-way ANOVA with Bonferroni post hoc test, ***p=0.0006; WT vs. TRPV1 KO: **p=0.0013; WT vs. TLR2 KO: ***p=0.0006. The data are expressed as the mean±SEM. ns, not significant.
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