Figure 2.
Beclin1 regulates APPL1+ endosomes. (A) MDA-MB-231 cells expressing shGFP, shBECN1 or shATG5 were stimulated with IGF-1 (100ng/ml) for the time periods indicated. Cells were fixed and stained with antibodies that recognize APPL1 (green). Nuclei were stained with DAPI (blue). Scale bar, 10 μm. (B) Quantification of APPL+ endosomes shown in (A). The data shown represent the mean of 23-37 independent cells. (C) Autophagic flux in MDA-MB-231 cells expressing shGFP, shATG5 or shBECN1. Cells were incubated for 4 hours in either complete or serum and amino acid-free media with or without 25 μM Chloroquine diphosphate. FBS, fetal bovine serum; AA, amino acids. *, p < 0.05; ***, p < 0.0005. See also Figure S2.