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. 2020 Jun 16;117(26):15281–15292. doi: 10.1073/pnas.2000500117

Fig. 8.

Fig. 8.

Effects of nanoparticle-encapsulated DOPr ligands on nociceptors. (A) Uptake of LipoMSN-Alexa647 (control) or DADLE-LipoMSN-Alexa647 into primary cultures of DRG neurons from DOPr-eGFP mice. Neurons were incubated with nanoparticles for 60 min. Representative images from two experiments, from four mice. (B and C) Rheobase of mouse DRG neurons at 0, 90, 120, or 180 min after exposure to DADLE, DADLE-LipoMSN-DADLE, DADLE-LipoMSN (all 100 nM), LipoMSN (control), or vehicle (control) and washing. Some neurons were exposed to PS2 and DADLE-LipoMSN-DADLE. Data points indicate the number of studied neurons from n = 6 to 12 mice in B and C for each treatment. Compared with *DADLE, ^DADLE-LipoMSN-DADLE, and #DADLE-LipoMSN; *#P < 0.05, **^^P < 0.01, ***P < 0.001, one-way (B) or two-way (C) ANOVA with Tukey’s post hoc test. (D) Colonic afferent activity at 0, 60, or 120 min after exposure of tissues to DADLE-LipoMSN-DADLE (100 nM). Some preparations were exposed to PS2 and DADLE-LipoMSN-DADLE. n = 5 mice per group. *P < 0.05, **P < 0.01, two-way (*) ANOVA with Sidak’s post hoc test. (E) Ipsilateral paw withdrawal responses in mice. DADLE, DADLE-LipoMSN-DADLE (both 100 nM DADLE), LipoMSN, or vehicle (Veh) was injected intrathecally at 48 h after intraplantar CFA. n = 5 mice per group. **P < 0.01, ****P < 0.0001 DADLE compared with DADLE-LipoMSN-DADLE, two-way ANOVA with Tukey’s multiple comparison post hoc test. (F) Uptake of LipoMSN-Alexa647 into endosomes of HEK293 cells expressing Rab5a-GFP after 120 min. (G) Time course of uptake of LipoMSN-Alexa647 into HEK293 cells. n = 3 independent experiments. (H) Rheobase of mouse DRG neurons. Neurons were incubated with LipoMSN-SDM25N (100 nM) or LipoMSN (control) for 120 min, washed (W), incubated with DADLE (10 nM, 15 min), and washed again. Rheobase was measured at 0 or 30 min after washing. Data points indicate the number of studied neurons from four mice for each treatment. *P < 0.05, **P < 0.01, two-way ANOVA with Tukey’s post hoc test. All results are mean ± SEM.