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. 2022 Jun 22;27(10):4009–4022. doi: 10.1038/s41380-022-01662-z

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© The Author(s) 2022

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. 2 — A Timeline of AAV injection, behavioural tests and location of AAV expression. B–E Inhibition of NICD in the mPFC suppressed METH-induced locomotor activity. B NICD mRNA (t₁₀ = 2.79, *P < 0.05) and protein (t₁₀ = 2.84, *P < 0.05) levels were significantly lower in syn-NICD-shRNA group as indicated by student’s t test. C, G Representative paths of mice in the syn-NICD-shRNA-METH and control-METH or syn-NICD-OE-METH and control-flag-METH groups on day 1 and day 23. D Downregulation of NICD in mPFC neurons significantly decreased locomotion on day 1 and day 23. Mixed-design ANOVA with a LSD post hoc multiple comparison revealed significant main effects of AAV [F _{(1, 20)} = 8.95, P < 0.01]_; METH [F _{(1, 20)} = 323.81, P < 0.001]; and AAV_×METH [F _{(1, 20)} = 8.10, P < 0.05]. E Time spent in the centre zone was significantly higher in the syn-NICD-shRNA-METH group than in the control-METH group on day 1 and day 23 [Mixed-design ANOVA, main effect of AAV, F _{(1, 21)} = 7.61, P < 0.05; METH, F _{(1, 21)} = 0.01, P > 0.05_; AAV × METH, F _{(1, 21)} = 7.18, P < 0.05]. F–I Overexpression of Notch1 in the mPFC enhanced METH-induced locomotor activity. F NICD mRNA (t₁₀ = −4.25, **P < 0.01) and protein (t₁₀ = −2.81, *P < 0.05) levels were significantly higher in the mPFC of the syn-NICD-OE group than the control-flag group as determined by student’s t test. H Overexpression of NICD significantly increased locomotion on day 1 and day 23 compared with the control-flag-METH group. Mixed-design ANOVA with a LSD post hoc multiple comparison revealed significant main effects of AAV [F _{(1, 20)} = 8.28, P < 0.01]; METH [F _{(1, 20)} = 241.37, P < 0.001]; but not the AAV×METH [F _{(1, 20)} = 2.70, P > 0.05_]. I The time that mice spent in the centre zone was significantly downregulated in the syn-NICD-OE-METH group compared with the control-flag-METH group on day 1 and day 23 [Mixed-design ANOVA, main effect of AAV, F _{(1, 22)} = 6.50, P < 0.05; METH, F _{(1, 22)} = 32.14, P < 0.001; AAV × METH, F _{(1, 22)} = 2.09, P > 0.05]. *P < 0.05, **P < 0.01, ***P < 0.001 vs. paired saline group; ^P < 0.05 vs. paired METH group; #P < 0.05, ###P < 0.001 vs. same group on day 1. Data were presented as mean ± S.E.M, n = 6–8.