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. 2013 Mar 27;3:1561. doi: 10.1038/srep01561

Figure 2. TAK1 overexpression induces autophagy in several cell types.

Figure 2

(A) GFP-LC3 was co-transfected with a control plasmid or a TAK1 plasmid. After transfection, the cells were incubated for 48 h, and GFP-LC3 puncta were then detected using fluorescence microscopy. Scale bars, 10 μm. (B) Cells were co-transfected with GFP-LC3 along with the indicated constructs for 48 h and then harvested for immunoblot analysis. β-actin was used as a loading control. GFP-LC3-II was detected as the faster migrating form of LC3 by immunoblot analysis on a 12% SDS-PAGE gel. The expression of TAK1, which was Flag-tagged, was measured using an anti-Flag antibody. (C) Cells were transfected with a control plasmid or a TAK1 plasmid for 48 h and then harvested for immunoblot analysis. Endogenous LC3 processing was evaluated by immunoblotting. (D) HEK 293T cells were co-transfected with GFP-LC3 and either TAK1-WT or TAK1-KW plasmid for 48 h. After transfection, cells were harvested and analyzed by immunoblottng.