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. 2022 Sep 21;19(4):1277–1292. doi: 10.1080/15548627.2022.2124500

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© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.

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Figure 5. — NEFM is present in AP and AL compartments of mouse cortical neurons. Cortices were double immunolabeled with NEFM and MAP1LC3 (A, unmodified images) or NEFM and CTSD antibodies (B, unmodified images). Images in C and D are modified in Photoshop (see Materials and Methods for details) to visualize NEFM-label in vesicular structures that is colocalized with MAP1LC3 (an AP marker, C, see arrows for colocalization) and CTSD (an AL/LY marker, Rudy-4, D, see arrows for colocalization) puncta. Colocalized puncta (yellow) were imaged through Z-stacks to show NEFM in the lumen of APs (see arrows in Ci to follow the same vesicles) and AL vesicles (see arrows in Di to follow the same vesicles) in mouse cortex layer III. Scale bars A-D, Ci and Di: 2 µm. Quantification of NEFM label in vesicular structures indicates 1/5 of NEFM label is colocalized with MAP1LC3 (E, n = 93 neurons), and CTSD (F, n = 109 neurons) vesicles in cortical neuronal cell bodies.