Fig. 4.
PAS antibody-immunoreactive proteins in mitochondria. SH-SY5Y cells were pre-incubated without and with wortmannin (40 nm, 30 min) prior to IGF-1 treatment (50 ng/mL, 15 min). (a) Purified cytosolic and mitochondrial proteins were separated in a 4–15% gradient SDS-polyacrylamide gel and immunoblotted with a phospho-Akt substrate (PAS) antibody. (b) Mitochondrial extracts from control, IGF-1, and wortmannin plus IGF-1 treated cells were immunoprecipitated with the PAS antibody and immunoblotted with ATP synthase β-subunit antibody and mitochondrial extracts were also immunoblotted for total levels of ATP synthase β-subunit.