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. 2020 Apr 20;295(23):8096–8105. doi: 10.1074/jbc.AC120.012595

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© 2020 Melick et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 3. — AKAP8L interacts with mTORC1 in the cytoplasm. A, AKAP8L localizes to the cytoplasm and nucleus. Expression of FLAG-tagged AKAP8L in mouse embryonic fibroblast cells. Red, FLAG-tagged AKAP8L (1.0 μg). B, expression of FLAG-tagged AKAP8L in the cytoplasmic and nuclear fractions of HEK293A cells. C, expression of endogenous AKAP8L in the cytoplasmic and nuclear fractions of HEK293A cells. D, Raptor interacts with AKAP8L in the cytoplasm. Co-immunoprecipitation of endogenous AKAP8L with HA-tagged Raptor. E, AKAP8L interacts with RIα. Co-immunoprecipitation of mCherry-tagged RIα with FLAG-tagged AKAP8L. F, Raptor interacts with AKAP8L and PKA machinery. Co-immunoprecipitation of FLAG-tagged AKAP8L, mCherry-tagged RIα, and FLAG-tagged PKA Catα with HA-tagged Raptor. IP, immunoprecipitation. s.e., short exposure; l.e., long exposure.