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. 2017 Jul 25;8:122. doi: 10.1038/s41467-017-00172-9

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© The Author(s) 2017

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 — Spindle matrix proteins regulate NSC reactivation and function downstream of insulin pathway during NSC reactivation. a NSC MARCM clones in control and Mtor ^k03905 were labelled with Mira, CD8 and Topro-3 (for DNA). b EdU, CD8 and Dpn were labelled in control and Mtor ^k03905 NSC clones. c Mira and Topro-3 or Dpn and EdU were labeled in NSCs from control and east RNAi knockdown. d, e Dpn and EdU or Mira were labelled in the VNC of control and east RNAi knockdown. f Larval brains from overexpression of an active form of InR (OE InRCA) with UAS-CD8-GFP, overexpression of InRCA with chro RNAi knockdown (OE InRCA + chro RNAi), overexpression of RHEB (OE RHEB) with UAS-CD8-GFP, and overexpression of RHEB with chro RNAi knockdown (OE RHEB + chro RNAi) were raised on sucrose-only (amino-acid free) food for 72 h and labelled by EdU. g Quantification of EdU⁺ cells per brain hemisphere in various genotypes. h Mira and Dpn were labelled in OE InRCA with UAS-CD8-GFP, OE InRCA with chro RNAi, OE RHEB with UAS-CD8-GFP and OE RHEB with chro RNAi on sucrose-only food. The right panels are enlarged views of the yellow boxes in the left panels. i Quantification of NSCs with a cellular extension in various genotypes. j Larval brains from OE InRCA with UAS-CD8-GFP and OE InRCA + chro RNAi were raised on sucrose-only food for 24 h and labelled by EdU. k Quantification of EdU⁺ cells per brain hemisphere for j. l Larval brains from PI3K (PI3K^CAAX) overexpression with UAS-CD8 or chro RNAi knockdown were raised on sucrose-only food for 72 h and labelled by EdU. All error bars indicate ± SD in g, i and k. *** indicates P < 0.001 in g, i and k by Student’s t-test. NSC clones were outlined by white dotted lines in a and b. Larval brain hemispheres were outlined by white dotted lines in f, h, j, l. Arrows indicated NSCs in b and arrowheads indicated the cellular extension of NSCs in a, c, e and h. Scale bars: 10 µm (a–c, f, h, j, l) and 20 µm (d, e)