Fig. 1

Experimental protocol for RCS and assessing pain behavior. a Schematic illustration of the RCS protocol. Mice were first kept at 7 °C in the RCS chamber from 7:00 pm to 10:00 pm (Before RCS). The mice were subjected to temperature changes between 7 °C and 22 °C every 30 min from 10:00 am to 5:30 pm, and were then maintained at 7 °C from 5:30 pm to 10:00 am for 7 days (RCS). After 7 days of RCS exposure, the RCS schedule ended at 10:00 am (After RCS). b Hind PWT was measured in the left and right hindlimbs using the electronic von Frey test. The PWT of the RCS group remained significantly lower than that of the untreated control (CTR) group in both the left and right hindlimbs until day 17 after RCS. n = 6 mice for each group. *p < 0.001, **p < 0.0001. Error bars indicate the SEM. Two-way ANOVA with Bonferroni's multiple comparisons test