Skip to main content
. 2014 Mar 4;19(3):431–444. doi: 10.1016/j.cmet.2014.02.010

Figure 2.

Figure 2

Mitochondrial AcCoA Production by Ach1p- or Mpc1p-Associated Pathways Is Required for Autophagy during Aging

(A) Fluorescence microscopy of GFP-Atg8p expressing wild-type (WT) and ACH1-deleted (Δach1) yeast grown to day 1 (young) and aged for 3 days in SC 2% glucose medium; propidium iodide (PI) counterstaining served to visualize dead cells. Scale bars represent 5 μm.

(B) Quantification of cells depicted in (A) with 150–300 counts (blinded) for each replicate. Autophagic cells were defined as cells with clear vacuolar GFP fluorescence. Data represent means ± SEM (n = 4). ∗∗∗p < 0.001.

(C) Representative immunoblot analysis of cells shown in (A) and further aged to day 5 using anti-GFP and anti-GAPDH (loading control) antibodies to detect “free-GFP” indicative of autophagic flux (see also Figure S2).

(D–F) Representative micrographs (D), respective quantification (E), and immunoblot analysis (F) of wild-type (WT) and MPC1 deleted (Δmpc1) yeast expressing GFP-Atg8p chimera as in (A)–(C) and aged to indicated time points. Data represent means ± SEM (n = 4). ∗∗∗p < 0.001.