Nature - USA (2020-09-24)

Extended Data Fig. 5 | The mTORC1 substrate-recruitment mechanism of
TFEB is determined by its N-terminal region. HeLa cells were transiently
transfected with plasmids expressing either GFP alone or GFP-tagged versions
of the following proteins: TFEB (GFP–WT–TFEB), a TFEB deletion mutant that
lacks the first 30 amino acids (GFP–Δ30TFEB), a chimeric protein in which the
first 30 amino acids of S6K (containing the TOS motif ) were fused to the
D30TFEB mutant (GFP-TOS-Δ30TFEB), or the TOS-D30TFEB chimeric protein in
which a key phenylalanine residue (F5) of the TOS motif was mutagenized to
alanine (GFP-F5A-Δ30TFEB). Twenty-four hours after transfection cell lysates
were incubated with GFP beads and subjected to immunoblotting using the
indicated antibodies (replicated three times).