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. 2015 Sep 22;11(11):2014–2032. doi: 10.1080/15548627.2015.1096485

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© 2015 The Author(s). Published with license by Taylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 3. — Defective autophagy in VSMCs elicits cellular hypertrophy, and increases migration capacity and total collagen amount. (A) VSMCs isolated from Atg7^+/+Tagln-Cre⁺ (+/+) and Atg7^F/FTagln-Cre⁺ (−/−) aorta were labeled with calcein AM and visualized by confocal fluorescence microscopy. Scale bar: 10 µm. Cell size was measured using z-stack images (*, P < 0.05; n = 2 experiments; Student t test). (B) Thoracic aorta of Atg7^+/+ and atg7^−/− mice were stained with H&E to measure the width of the media (white arrows) (**, P < 0.01; n = 6 regions/aorta; Univariate). Scale bar: 25 µm. Note that the number of VSMC layers between Atg7^+/+ and atg7^−/− aorta was not different. (C) Migratory capacity of Atg7^+/+ and atg7^−/− VSMCs was analyzed using an Oris Migration Assay (***, P < 0.001; n = 2 experiments in triplicate; Student t test). Western blot analysis of TGFB and CXCL12 in Atg7^+/+ and atg7^−/− VSMCs. (D) Atg7^+/+ and atg7^−/− VSMCs were left untreated or treated with 10 ng/ml TGFB for 48 h and stained with Sirius red to examine total collagen amount (***, P < 0.001 vs. Atg7^+/+; ^###, P < 0.001 vs. control; n = 4 experiments in triplicate; two-way ANOVA with genotype and treatment as category factors).