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. 2020 Apr 3;11:1674. doi: 10.1038/s41467-020-15412-8

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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. 3 — a Celf1 (red) and Pax6 (green) VZ, IHC at E13 and E16 (n = 3 animals per age). DAPI shown in blue. b Western blot of Celf1 protein (upper panel) expression in VZ and CP from LCM neocortices at E13 and E16 (n = 3 animals per age). The Celf1 upper band is putatively Celf1 Long (Celf1L); the Celf1 lower band is putatively Celf1 Short (Celf1S), indicated at left. Gapdh was used as a loading control (bottom panel). c Densitometry quantification of b. Data represent the mean and SEM. Data normalized to Gapdh and then to E13. Statistics: Student’s t-test. *p < 0.05. d Celf1 (red, top) or Elavl4 (red, bottom) IHC on Nestin-GFP (green) neocortices at E13 and E16 (n = 4 animals per age). e RNA immunoprecipitation (RIP) from E13 and E16 neocortices. Relative mRNA levels were determined by qRT-PCR from Celf1 vs. IgG RIPs (n = 4 separate RIPs, 6 neocortices per age). Pan-Elavl4 and specific Elavl4 isoforms are shown in addition to ActB and Gapdh as non‐binding controls. f RIP from N2a cells (n = 3 RIPs from separate transfections). Data represent the mean and SEM. Data normalized to Gapdh. Celf1 was used as positive control⁶⁰. 18S was used as negative control. Statistics: Two-way ANOVA with Tukeyʼs. ****p < 0.0001.