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. 2021 Apr 14;11:220. doi: 10.1038/s41398-021-01337-3

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Panel (a) depicts the experimental timeline. Adult male Long-Evans rats were first trained to drink alcohol for 4 weeks in the intermittent access 2-bottle free-choice (IA2BC) paradigm, followed by training for ethanol self-administration (SA) in the operant chambers in the FR1 paradigm for 8–10 weeks. They then were exposed to either CIE or Air vapor for 8 weeks. The number of active lever presses (b) and ethanol consumption (c) of CIE exposed rats significantly increased compared to before CIE or Air control rats. At 24-h abstinence from the last drinking session, a selective FAAH inhibitor (URB597), MAGL inhibitor (JZL184), CB1R agonist WIN55,212-2 (WIN), or Vehicle was bilaterally injected into the LHb 30 min before a drinking session. Panel (d–l) summarizes the effect of these compounds on active lever presses in both Air (d, g, and j) and CIE rats (e, h, and k), as well as the change of ethanol consumption (f, l). All data are expressed as mean ± SEM. ^^^p < 0.001 vs baseline (BL) within CIE or Air group revealed by one-way RM ANOVA, ***p < 0.001 vs Vehicle or URB597/JZL184/WIN + RIM within CIE group revealed by two-way ANOVA; ^#p < 0.05 vs Vehicle or URB597/JZL184/WIN + RIM within CIE group, by one-way ANOVA followed by Bonferroni post-hoc test. n = 16-18 rats/group.