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. 2020 Jan 31;11:645. doi: 10.1038/s41467-019-14164-4

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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 Still images from an intravital imaging time-lapse movie of liver from KFERQ-Dendra mice showing the dynamics of several CMA lysosomes (arrows). Similar results were obtained in four independent experiments. b Representative two-photon microscopy images of livers from fed or 24 h starved KEFRQ-Dendra and KFERQ-Dendra-L2A^null mice. Insets: higher magnification. Similar results were obtained in three independent experiments. c, d Representative images of liver and kidney (c) and WAT and BAT (d) from 24 h starved KFERQ-Dendra mice. In c mice were injected with 155 KDa TRITC Dextran (red) to highlight lysosomal compartments. Similar results were obtained in four independent experiments, Insets: Boxed area at higher magnification. e Top: Maximum projection of cortical (left) and hippocampus (right) regions from KFERQ-Dendra mice imaged with two-photon microscopy. Bottom: examples of magnified individual cells. Similar results were obtained in four independent experiments. f Left top: KFERQ-Dendra mice liver section with delineated sub-zoning. CV: central vein, L1 PC: layer 1 pericentral, L5 PP: layer 5 periportal. DAPI is shown in blue. Right: Higher magnification liver region. Bottom: LAMP1 staining of full field (left) and higher magnification region with delineated zones (right) of liver. Right: Number of KFERQ-Dendra (top) and LAMP1 (bottom) positive puncta in liver regions. Values are from 16 (Dendra) and 12 (LAMP1) sections coming from 4 independent experiments. g Image of livers from KFERQ-Dendra mice fed, starved for 24 h or paraquat (PQ) injected. Insets: higher magnification of regions distal from the CV. Yellow arrow: KFERQ-Dendra positive puncta gradation. h Fluorescent puncta per cell in regions of starved or PQ-treated KFERQ-Dendra mice livers. i Overall number of fluorescent puncta in the same livers as (g). Values in h and i are from 11 (CV), 12 (L1PC) and 12 (L5PP) sections (panel h), and 16 (fed), 11 (starved) and 11 (PQ) sections (panel i) Data are presented as mean values ± s.e.m. One-way ANOVA test followed by Dunnett’s multiple comparisons test was used. Source data are provided in the Source Data file.