Fig. 8.
Contribution of Dnm2 to PAR2-evoked inflammation and pain in the colon. (A–H) Dnm2 or control (Ctrl) shRNA was administered to Par2-mugfp mice by intracolonic injection. Mice were studied after 48 h. (A) Localization of Dnm2 by RNAScope. Representative images are shown. (Scale bars, 100 µm.) (B) Quantification of Dnm2 in the colon. (C) Localization of GFP immunoreactivity in the colon at 5 h after intracolonic injection of 2F. Arrows denote endosomes; arrowheads denote cell surface. (Scale bars, 10 µm.) (A–C) n = 5 mice. (D–F) Levels of TNF-α (D), CXCL1 (E), and IL-1β (F) in the colon 5 h after intracolonic injection of 2F. (G and H) Abdominal von Frey filament withdrawal responses 1–5 h after intracolonic injection of 2F. (G) Time course. (H) Area under the curve (AUC) from 0 to 5 h. (D–H) n = 6 mice. (I and J) Abdominal von Frey filament withdrawal responses of wild-type, Par2-mugfp, and Par2Nav1.8 mice 1–6 h after intracolonic injection of 2F. (I) Time course. (J) Area under curve from 0 to 6 h. n = 5 mice. (K) Effects of 2F (10 µM) on IL-8 release from explants of human colonic mucosa. Explants were preincubated with dyngo4a or vehicle (DMSO, control). Mean ± SEM; n = 6. (B) **P < 0.01 by Student’s t test. (D–J) *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by one- and two-way ANOVA or Tukey’s test. (K) **P < 0.01 by Student’s t test.