Non-invasive dural application of IS produces headache-related behavior in both male and female mice. Dural application of IS produces cutaneous facial hypersensitivity in both male (a) and female mice (d). Mice treated with IS (1–24 hr; n = 6–10), showed significant facial (a) and (c) responses, but no significant differences in hindpaw sensitivity were observed ((b) and (d)). Significant differences among means for each group were determined by two-way ANOVA followed by Bonferroni post hoc test. Males facial: Time F (4, 70) = 1.95, p= 0.1117, treatment F (1, 70) = 18.54, p < 0.0001; hindpaw: Time F (4, 55) = 2.903, p= 0.0299, treatment F (1, 55) = 1.128, p= 0.2928. Females facial: Time F (4, 75) = 5.908, p= 0.0003, treatment F (1, 75) = 21.28, p < 0.0001; hindpaw: Time F (4, 60) = 1.111, p= 0.3599, treatment F (1, 60) = 9.261, p= 0.0035.