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. 2017 Dec 17;13(12):2041–2055. doi: 10.1080/15548627.2017.1376160

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© 2017 Max Planck Institute for Infection Biology

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 2. — NEDD4 contributes to autophagy. Western blot analysis of LC3 levels in NEDD4 Scr and KD HeLa cells (A), NEDD4 Scr and KD THP-1 cells (B) and Nedd4^+/+ or nedd4^−/− MEFs (C) with or without choloroquine (CQ) treatment. (D) Western blot analysis of SQSTM1 levels in Nedd4^+/+ or nedd4^−/− MEFs with or without CQ treatment. (E) Immunofluorescence analysis of YFP-LC3B puncta in Nedd4^+/+ or nedd4^−/− MEFs stably expressing YFP-LC3B. (F) Quantification of YFP-LC3B puncta per cell. Data are shown as mean ± SD of 3 independent experiments. A total of 50 cells per group for each experiment were quantified. P values were calculated using the Student 2-tailed unpaired t test. (*) p ≤0.05. (G) Western blot analysis of LC3 levels in Nedd4^+/+ or nedd4^−/− MEFs after starvation, Torin1 or CCCP treatment with or without CQ. (H) Western blot analysis of LC3 levels in Scr and NEDD4 KD THP-1 cells upon Mtb or Mtb ΔRD1 infection. (I) Western blot analysis of LC3 levels in Scr and NEDD4 KD THP-1 cells upon BCG or rBCG infection. LC3-II:ACTB ratio was determined using ImageJ software and normalized to control. Results are representative of 3 independent experiments.