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. 2021 Oct 9;297(5):101291. doi: 10.1016/j.jbc.2021.101291

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© 2021 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

DEPTOR is phosphorylated on tyrosine 289.A, denaturing FLAG immunoprecipitation of 293T cells transfected with FLAG-YFP (negative control) or FLAG-DEPTOR. DEPTOR tyrosine phosphorylation was detected using antiphosphotyrosine antibodies (4G10). B, schematic representation of the different DEPTOR fragments. C, 293T cells transfected with FLAG-YFP (negative control), FLAG-DEPTOR^WT, FLAG-DEPTOR N-terminal region, or FLAG-DEPTOR C-terminal region were lysed for immunoprecipitation in denaturing condition. Immunoprecipitated DEPTOR phosphorylation was assessed using antiphosphotyrosine antibody 4G10. D, 293T cells transfected by FLAG-YFP (negative control), FLAG-DEPTOR C-terminal region, FLAG-DEPTOR D3, and FLAG-DEPTOR D4 were lysed for immunoprecipitation in denaturing condition. Immunoprecipitated DEPTOR phosphorylation was assessed using antiphosphotyrosine antibody 4G10. E, DEPTOR tyrosine phosphorylation is located on the residue 289. 293T cells were transfected by FLAG-YFP (negative control), FLAG-DEPTOR-D4^WT, FLAG-DEPTOR-D4^Y289F, FLAG-DEPTOR-D4^Y326F, or FLAG-DEPTOR-D4^Y289-326F, and a denaturing immunoprecipitation was made to evaluate their phosphorylation status.