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. Author manuscript; available in PMC: 2010 May 1.
Published in final edited form as: Autophagy. 2009 May 6;5(4):520–526. doi: 10.4161/auto.5.4.7768

Figure 6.

Figure 6

Autophagy is upregulated in response to chemical inducers functioning independently of the TOR pathway. (A) GFP-Lc3 embryos at 3 dpf were treated with the indicated drugs or DMSO for 24 h, and then stained with LysoTracker Red for 1 h before imaging by confocal fluorescence microscopy. (B) Colocalization of GFP-Lc3 dots with LysoTracker Red (LT) in (A) was quantified using Leica Simulator SP5 software. (C and D) Wild-type (C) or GFP-Lc3 (D) embryos at 2 dpf were treated with the indicated chemicals or the solvent DMSO (—) for 24 h. Protein extracts were analyzed by SDS-PAGE and detected by western blot using anti-LC3, or anti-tubulin antibody as a loading control. (C, right) The relative increase of the Lc3-II/Lc3-I ratio induced by chemicals was quantified by ImageJ software (http://rsb.info.nih.gov/ij/) and represented as mean ± s.d. of three independent experiments. The value of Lc3-II/Lc3-I from the DMSO-treated sample was set to 1.0 and other values were normalized. Rap, rapamycin; Cal, calpeptin; ddA, 2’5’-dideoxyadenosine; Ver, verapamil; clon, clonidine.