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. 2008 Aug 25;182(4):685–701. doi: 10.1083/jcb.200803137

Figure 2.

Figure 2.

Starvation-induced and PI(3)P-dependent translocation of DFCP1 to punctate structures partially colocalizing with autophagosomes. (A) Lysates from cells expressing WT DFCP1 or the DM were incubated with Affigel beads coupled to PI(3)P; WT DFCP1 bound to PI(3)P, whereas the DM did not. in, 5% of input. (B) Stable HEK-293 cell lines expressing WT DFCP1 or the DM were left untreated or starved of amino acids for 90 min. Cells were then fixed and stained for DFCP1. Both WT and DM DFCP1 under normal conditions localized to the ER/Golgi; after starvation, only the WT translocated to punctate structures. (C) HEK-293 cells stably expressing WT DFCP1 were starved alone or in the presence of 3-methyladenine (MA) or wortmannin (wortm) as indicated before fixation and staining for DFCP1. (D) Starved HEK-293 cells expressing DFCP1 were costained with markers to the ER such as calnexin and KDEL. (E) HEK-293 cells stably expressing WT DFCP1 were transfected with GFP-MAP-LC3 and starved for 90 min as indicated. Note that in cells expressing WT DFCP1, both MAP-LC3 and DFCP1 translocated to punctate structures upon starvation, some of which colocalized. Bars: (A–C and E) 20 μm; (D) 0.5 μm.