AMDE-1 Is a Dual Function Chemical for Autophagy Activation and Inhibition (original) (raw)
Fig 3
AMDE-1 can inhibit autophagic degradation.
(A-B) MEFs were treated with or without AMDE-1 (10 μM) and CQ (40 μM) for 20 h. Endogenous LC3 was analyzed by immunoblot assay (A) and densitometry (B). (C-D) MEFs were treated with AMDE-1 (10 μM) for 0–20 h as indicated. The level of SQSTM1/p62 was analyzed (C) and quantified (D). (E-F) HEK-293A cells expressing GFP-RFP-LC3 were treated with CQ (40 μM), Rap (1 μM) and AMDE-1 (10 μM) for 6 h (E). Puncta showing both green and red fluorescence (indicated as yellow), or showing only the red fluorescence (indicated as red) were quantified (F). (G-H) MEFs were incubated in complete medium (CM), or in EBSS with or without AMDE-1 (10 μM), 3-MA (10 mM), CQ (10 μM), Rap (1 μM) or CBZ (50 μM) for 6 h (G) or 16 h (H). The long-lived protein degradation was measured. For panel B, D, F, G and H, values represent means ± SD from three independent experiments. ***: p<0.001; **: p<0.01.