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. Author manuscript; available in PMC: 2015 Jan 29.
Published in final edited form as: J Cell Physiol. 2014 Oct;229(10):1359–1368. doi: 10.1002/jcp.24569

Figure 5. CMA regulates the unphosphorylated form of PED.

Figure 5

(A) Schematic representation of PED phosphorylation sites and putative KFEQR sites. (B) HEK-293 were transfected with PED-Myc cDNA, HA-PED-S104/116A and HA-PED-S104/116D. After 48 hrs, 1 mg of protein extracts were immunoprecipitated with anti-PED antibody or with control IgG. Samples were then resolved with SDS-PAGE and western blot with anti-Hsc70 antibodies. 10 μg of total cellular extract was loaded as control. (C) A549 cells transfected with PED-Myc, HA-PED-S104/116A and HA-PED-S104/116D were serum deprived for 24 hrs. Protein levels of PED wt and phosphorylation mutant were analyzed by Western blot (D) Western blot analysis of PED-Myc and HA-PED-S104/116A, HA-PED-S104/116D after A549 co-transfection with LAMP2A cDNA upon 24 hr. (E) Western blot analysis of PED-Myc and HA-PED-S104/116A, HA-PED-S104/116D upon A549 treatment with 15 mMol of NH4Cl for 4 hr. (F) Western blot analysis of PED-Myc and HA-PED-S104/116A, HA-PED-S104/116D upon A549 treatment with 10 μMol of chloroquine for 24 hr.