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. 2023 Oct 13;49(2):443–454. doi: 10.1038/s41386-023-01739-5

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — *An example week of the variable-dose CNO treat paradigm is shown in Table S1. A 100% of male (n = 7) and female (n = 8) DREADD+ mice continued consuming daily CNO treats across 9 weeks. B Weekly body weight measurements across 9 weeks in DREADD- (n = 10) and DREADD+ (n = 7) males (2-way RM ANOVA; F_time(1.091,16.36) = 5.050, p = 0.036, F_genotype(1,15) = 0.043, p = .838; F_{time*genotype}(9,135) = 0.162, p = 0.997). C Lower, variable dose CNO treats did not induce a greater overall body weight change in DREADD+ males compared to controls (unpaired t-test; t₍₁₅₎₌0.434, p = 0.670). D Weekly body weight measurements across 9 weeks in DREADD- (n = 8) and DREADD+ (n = 8) females (2-way RM ANOVA; F_time(1.230,17.22) = 2.238, p = 0.15; F_genotype(1,14) = 0.039, p = 0.847; F_{time*genotype}(9,126) = 0.151, p = 0.998). E There were no differences in overall body weight change at the end of 9 weeks in DREADD+ females compared to controls (unpaired t-test; t₍₁₄₎₌0.684, p = 0.505; n = 8 per group). F 9 weeks of CNO did not affect HPA axis reactivity to an acute CNO injection (2-way RM ANOVA; F_{CNO chronicity}(1,9) = 0.112, p = 0.746; F_time (1.564, 14.08) = 13.09, p = 0.001; F_{time*CNO chronicity}(3, 27) = 0.494, p = 0.690) or (G) total amount of corticosterone released following an acute CNO injection in DREADD+ males (n = 7) compared to CNO-naïve males (n = 3) (unpaired t-test; t₍₈₎ = 0.0467, p = 0.964). H 9 weeks of chronic CNO did not affect HPA axis reactivity to an acute CNO injection (2-way RM ANOVA; F_{CNO chronicity}(1,10) = 0.002, p = 0.967; F_time(1.686, 16.86) = 40.77, p < 0.0001; F_{time*CNO chronicity}(3, 30) = 0.379, p = 0.769) or (I) total amount of corticosterone released in female DREADD+ mice (n = 6) compared to CNO-naïve females (n = 6) (unpaired t-test; t₍₁₀₎ = 0.221, p = 0.829). J Slope analysis of HPA axis corticosterone response from 60- to 120- mins post-injection. Female DREADD+ mice recovered faster from peak corticosterone levels than DREADD+ males when CNO-naïve (p = 0.014) and after 9 weeks of chronic CNO administration (p = 0.003) (2-way ANOVA; F_sex(1,18) = 18.49, p = 0.0004; F_{CNO chronicity}(1,18) = 1.231, p = 0.282; F_{sex*CNO chronicity}(1,18) < 0.0001, p = 0.999). (*p < 0.05, @ main effect of sex).