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. 2023 Mar 31;14:1800. doi: 10.1038/s41467-023-36808-2

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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 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. 6 — A Experimental timeline for DiD electrophysiology studies. B Average alcohol intake in DiD, two-way Student’s unpaired t-tests. C Cumulative alcohol intake in DiD, two-way Student’s unpaired t test. D Average alcohol preference in DiD, two-way Student’s unpaired t tests. E Resting membrane potential (RMP) in LHb_5HT2c neurons. F LHb_5HT2c rheobase. G LHb_5HT2c action potential (AP) threshold. H LHb_5HT2c elicited APs, females, two-way repeated measures ANOVA. I LHb_5HT2c elicited APs, males, two-way repeated measures ANOVA followed by Holm-Sidak’s post-hoc, p_DiD = 0.0236, p_DiDXcurrent = 0.0512. J Average RMP in BNST_5HT2c neurons, p_sex = 0.0190. K BNST_5HT2c rheobase. L BNST_5HT2c AP threshold. M BNST_5HT2c elicited APs, females, two-way repeated measures ANOVA. N BNST_5HT2c elicited APs, males, two-way repeated measures ANOVA. O Voltage–Current plot for LHb_5HT2c females, two-way repeated measures ANOVA. P Voltage–Current plot for LHb_5HT2c males, two-way repeated measures ANOVA. Q Voltage–current plot for BNST_5HT2c females, two-way repeated measures ANOVA, p_DiD = 0.1032, p_DiDXvoltage = 0.0701. R Voltage–Current plot for BNST_5HT2c males, two-way repeated measures ANOVA. S Representative trace in medial LHb_5HT2c during rheobase test. T Quantification of rheobase values in medial LHb_5HT2c in females, two-way Student’s unpaired t test, p = 0.0157 (water n = 5 mice/7 cells; DiD n = 5 mice/10 cells). U Breakdown of cell properties at RMP, BNST females. V Breakdown of cell properties at RMP, LHb females. W Breakdown of cell properties at RMP, BNST males. X Breakdown of cell properties at RMP, LHb males. Depol block = silent at greater than −40 mV. BNST Females: water n = 6 mice/10 cells, DiD n = 6 mice/19 cells; BNST Males: water n = 5 mice/16 cells, DiD n = 5 mice/16 cells. LHb Females: water n = 7 mice/14 cells, DiD n = 6 mice/19 cells; LHb Males: water n = 6 mice/19 cells, DiD n = 6 mice/19 cells. Unless otherwise stated, statistical comparisons were performed using two-way ANOVA followed by Holm-Sidak’s post-hoc. All data except for box plots are represented as mean ± SEM. For box plots, center line is the mean, box limits are 25th–75th percentiles, whiskers are min to max. & denotes effect of DiD, @ denotes effect of sex, # denotes interaction (sex × DiD or DiD × current), and * denotes post-hoc p < 0.05. Source data are provided as a Source Data file. Created with Biorender.com.