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. 2015 Jan 15;29(2):129–143. doi: 10.1101/gad.250282.114

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© 2015 Froldi et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 6. — Nerfin-1 is required for timely NB differentiation, and ectopic nerfin-1 promotes differentiation. (A–B″) At the end of neurogenesis (120 h ALH), Nerfin-1 and Pros are localized to the NB (Mira⁺, red) nucleus, prior to NB terminal differentiation. In 24% of NBs, both Pros (white) and Nerfin-1 (green) are expressed in the nucleus (A–A″), and 29% of NBs express only Nerfin-1 but not Pros (B–B″). (C) Histogram depicting the percentage of nuclear localization of Pros and Nerfin-1 in the NBs at 96 h (n = 20) and 120 h (n = 17). (D–D′) nerfin-1¹⁵⁹ NBs (Dpn⁺, white; yellow arrowheads) fail to undergo timely terminal differentiation at 120 h. In these NBs, nuclear Pros (red) localization does not occur. (E–M) Nerfin-1 overexpression in NBs with pan-NB driver dnab-Gal4 induces premature differentiation and reduction in the overall NB numbers at 96 h (E,F) and 110 h (G,H; maximum projections), as quantified in M (96 h, control: m = 122, SEM = 1.45, n = 3; dnab>nerfin-1: m = 82.4, SEM = 2.49, n = 10; 110 h, control: m = 125, SEM = 1.8, n = 3; dnab>nerfin-1: m = 58, SEM = 2.6, n = 5). (I,J) This reduction in NB numbers corresponds with a reduction in NB diameter, as quantified in L (control: m = 13.1 μm, SEM = 0.36 μm, n = 27; dnab>nerfin-1: m = 10.4 μm, SEM = 0.3 μm, n = 24), and increase in the incidence of size-symmetric divisions indicative of premature differentiation. (K) Histogram depicting the average diameter of thoracic NBs (Mira⁻) and their progeny (Mira⁺) during telophase (control, Mira⁻: m = 10.5 μm, SEM = 0.3 μm, n = 10; Mira⁺: m = 5.3 μm, SEM = 0.4 μm, n = 10; dnab>nerfin-1, Mira⁻: m = 5.8 μm, SEM = 0.5 μm, n = 8; Mira⁺: m = 4.6 μm, SEM = 0.6 μm, n = 8). (N) Schematic depicting the localization of Nerfin-1 during larval and pupal neurogenesis. In larval stages (up to 96 h ALH), Nerfin-1 is expressed in neurons (which also express the differentiation marker Pros) and not in self-renewing NBs or GMCs. At the end of neurogenesis (120 h ALH), Nerfin-1 and Pros are localized to the nucleus of the NBs prior to their terminal differentiation. Bars: E–H, 100 μm; all others, 10 μm. (****) P ≤ 0.0001. See also Supplemental Figures S16, S17.