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. 2017 Oct 25;7:13999. doi: 10.1038/s41598-017-14376-y

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© The Author(s) 2017

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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Increasing MCFA ratio protected HepG2 cells against LCFAs-induced fat accumulation, insulin resistance, and lipotoxicity by rescuing autophagy. To investigate the role of autophagy in MCFA-mediated protections, we blocked autophagy in MCF2-loaded HepG2 cells using either Bafilomycin A1 (5 nM) or chloroquine (10 μM). After treatment, fat accumulation (A), insulin resistance (B and C), ER stress, and apoptosis (D) were analyzed. Values are mean ± SEM (n = 3). For densitometric analyses of Western blotting data, β-actin or total Akt (for Akt phosphorylation analyses only) were used as the loading controls. *, ^#(vs. BSA treated cells without insulin stimulation), and ^†(vs. SDF treated cells without insulin stimulation) indicate statistical significance, P < 0.05. n.s.: no significant difference. SDF: standard fat loading mixture (0.4 mM PA + 0.4 mM OA); MCF2: MCFA-containing fat mixture 2 (0.4 mM PA + 0.2 mM OA + 0.2 mM LA); Baf: Bafilomycin A1. CQ: chloroquine. Full-length blots are presented in Supplementary Figure S4.