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    Nature
    https://www.nature.com › articles

    mTOR-dependent phosphorylation controls TFEB nuclear export

    • Cells were cultured in the following media: HeLa (ATCC) in MEM (Cat# ECB2071L, Euroclone); HEK293T (ATCC) in DMEM high glucose (Cat# ECM0728L, Euroclone); ARPE19 (ATCC) in DM… See more

    Materials

    Reagents used in this study were obtained from the following sources: Antibodies to Phospho … See more

    Nature
    Plasmids

    The plasmid-encoding ΔNLS-TFEB-GFP was a kind gift of Shawn Ferguson (Yale School of Medicine, CT, USA). TFEB S138A-GFP was a kind gift of Chonglin Yang (Chin… See more

    Nature
    Cell Treatments

    For experiments involving amino acid starvation, cell culture plates were rinsed twice with PBS and incubated in amino acid-free RPMI (Cat# R9010-01, USBiologic… See more

    Nature
    High-Content Analysis

    HeLa TFEB-GFP cells were seeded in 96-well plates and incubated for 24 h. After incubation cells were treated as described above, rinsed with PBS once, fixed for 10 min with 4… See more

    Nature
    Cell Lysis and Western Blotting

    Cells were rinsed once with PBS and lysed in ice-cold lysis buffer (250 mM NaCl, 1% Triton, 25 mM Hepes pH 7.4) supplemented with protease and phosphatas… See more

    Nature
    Does TFEB contain a CRM1 consensus hydrophobic sequence?
    We found that TFEB contains a CRM1 consensus hydrophobic sequence located at the N-terminal portion of the protein that is highly evolutionary conserved (Fig. 3a) and that is also present in other MiT-TFE members, such as TFE3 and MITF (Fig. 3b).
    nature.com