Figure 4

Location of CDK1 in the autophagosome/lysosome-enriched cellular fraction. (A) Subcellular fractions were prepared from MCF7 cells treated or not with concanamycin A (Con A) or bafilomycin A1 (Baf A1) for 4.5 hours, as described in Materials and Methods. Equal amounts of protein were analyzed by SDS-PAGE and immunoblot. The fractions were tested for purity against α Tubulin (cytosolic fraction), NBS1 (nuclear fraction), and LC3 (lysosome rich fraction). (B, C) MCF7 cells were pretreated with Con A for 30 min and then treated with MG132 (4 hours) (B) or etoposide (Eto, 24 hours) (C). Subcellular fractions prepared as in (A) were analyzed by Western blot.