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. 2015 Jan 28;10(12):2297–2309. doi: 10.4161/15548627.2014.994359

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© 2014 The Author(s). Published with license by Taylor & Francis

© Natalie Spang, Anne Feldmann, Heike Huesmann, Fazilet Bekbulat, Verena Schmitt, Christof Hiebel, Ingrid Koziollek-Drechsler, Albrecht M Clement, Bernd Moosmann, Jennifer Jung, Christian Behrends, Ivan Dikic, Andreas Kern, and Christian Behl

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 6. — The RAB3GAP complex antagonizes the FEZ1/2-mediated suppression of autophagy. (A) Immunoblot analysis of cells that were manipulated with the indicated siRNA for 48 h and treated with DMSO (−) or bafilomycin A₁ (+) for 4 h. Tubulin served as control for equal loading. Statistics are depicted as mean ± SD normalized to nonsense control; n.s. = not significant, *P < 0.05, **P < 0.01, n = 4, t test. (B) Identical to (A) but different siRNA treatments. Statistics are depicted as mean ± SD normalized to nonsense control; n.s. = not significant, *P < 0.05, **P < 0.01,n = 4, t test. (C) Confocal images of LC3 immunostainings. Fibroblasts were treated with the indicated siRNA for 48 h and treated with DMSO or bafilomycin A₁ for 4 h. DAPI was used to stain nuclei. Scale bar = 50 μm. Autophagosomal structures were counted in 20 to 40 cells of 3 independent experiments; n.s. = not significant, ***P < 0.001, t test. (D) Immunoblot analysis of cells that were manipulated with the indicated plasmids for 48 h and treated with DMSO (−) or bafilomycin A₁ (+) for 4 h. Since FEZ1/2 overexpression affected tubulin levels, actin served as control for equal loading. Statistics are depicted as mean ± SD normalized to eV control; n.s. = not significant, *P < 0.05,n = 4, t test.