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. 2020 Aug 26;11:4276. doi: 10.1038/s41467-020-17861-7

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© The Author(s) 2020

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. 4 — a Experimental design for t-LTP induction in two clustered spines (<5 µm) with PEP1-TGL (200 µM) inside the pipette. b Representative experiment where two clustered spines were activated with t-LTP pre–post pairing of +7 ms. Traces correspond to an average of 10 uEPSPs recorded in the soma and generated by 2P uncaging before (control, black trace) and after the induction of t-LTP with PEP1-TGL (red trace) over the indicated spines (red dots). c Time course of uEPSP amplitude, neck length, and spine head volume (P = 0.75, P < 0.001, P = 0.96, respectively, n = 5 spine pairs, one-way repeated-measures ANOVA) changes following STDP induction (+7 ms) in clustered spines with PEP1-TGL. n.s. not significant; **P < 0.01, post hoc Dunnet’s test. d Changes in uEPSP amplitude, neck length, and head volume of activated clustered spines 15–25 min after t-LTP induction (+7 ms) in control conditions (Cont) and with PEP1-TGL (PEP1-TGL: uEPSP = 94.82 ± 14.82%, P = 0.81; neck length = 79.71 ± 4.32%, P = 0.0078; spine head volume = 100.08 ± 3.23%, P = 1, n = 5 spine pairs, two-sided Wilcoxon test; *P < 0.05, **P < 0.01). e Experimental design for t-LTP induction in two clustered spines (<5 µm) with Latrunculin-A (Lat-A, 100 nM). f Same as b, but with Lat-A. g Time course of uEPSP amplitude, neck length, and spine head volume (P < 0.001, P = 0.35, P = 0.72, respectively, n = 9 spine pairs, one-way repeated-measures ANOVA) changes following STDP induction (+7 ms) in clustered spines with Lat-A. n.s. not significant; **P < 0.01, post hoc Dunnet’s test. h Changes in uEPSP amplitude, neck length, and head volume of the activated clustered spines 15–25 min after t-LTP induction (+7 ms) in control conditions (Cont) and with Lat-A (Lat-A: uEPSP = 69.55 ± 7.13%, P = 0.008; neck length = 93.04 ± 6.01%, P = 0.32; spine head volume = 99.80 ± 4.33%, P = 0.95, n = 9 spine pairs, two-sided Wilcoxon test; *P < 0.05, **P < 0.01). Shaded area and error bars represent SEM. Lines, bars, and dots in c, d, g, h: uEPSP = black, neck length (NL) = red, and head volume (HV) = blue.