Figure 5.
Figure 5. Early stage of autophagosome formation is suppressed in β−/− MEFs. (A) β+/+ and β−/− MEFs were transfected with GFP-DFCP1 expression construct. 24 h after transfection, cells were cultured in complete medium, serum-free medium (6 h), or Hank’s buffer (2 h). Cells were fixed, observed, and imaged under a deconvolution fluorescence microscope. Only cells with moderate levels of GFP signal were quantified and imaged. Representative images are shown. Bar, 20 µm. Quantification of GFP puncta per cell is shown on the right. Error bars = SEM; n = 10; *, P < 0.05; ***, P < 0.001; N.S., nonsignificant. (B) β+/+, β−/−, and p110-β–reconstituted β−/− MEFs lysates were probed for Atg5 and β-tubulin antibodies. The quantification of relative Atg5–Atg12 conjugation is shown on right. Data shown are the mean values from three independent experiments (*, P < 0.05; N.S., nonsignificant; error bars = SEM).

Early stage of autophagosome formation is suppressed in β−/− MEFs. (A) β+/+ and β−/− MEFs were transfected with GFP-DFCP1 expression construct. 24 h after transfection, cells were cultured in complete medium, serum-free medium (6 h), or Hank’s buffer (2 h). Cells were fixed, observed, and imaged under a deconvolution fluorescence microscope. Only cells with moderate levels of GFP signal were quantified and imaged. Representative images are shown. Bar, 20 µm. Quantification of GFP puncta per cell is shown on the right. Error bars = SEM; n = 10; *, P < 0.05; ***, P < 0.001; N.S., nonsignificant. (B) β+/+, β−/−, and p110-β–reconstituted β−/− MEFs lysates were probed for Atg5 and β-tubulin antibodies. The quantification of relative Atg5–Atg12 conjugation is shown on right. Data shown are the mean values from three independent experiments (*, P < 0.05; N.S., nonsignificant; error bars = SEM).

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