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. 2022 Feb 18;33(3):ar26. doi: 10.1091/mbc.E21-06-0309

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FIGURE 7: — The interaction of TFEB with mTOR, but not with PP2A, is disrupted by PIKfyve suppression. (A) Lysates from HEK293T cells transfected with FLAG-mTOR were immunoprecipitated with anti-FLAG antibodies and then analyzed by immunoblot using the indicated antibodies. (B) Quantitation of immunoprecipitated TFEB signal intensities from immunoblots in A and normalized to input TFEB protein (mean ± SD; three independent experiments). **P < 0.01. (C) Lysates from HEK293T cells transfected with TFEB-FLAG were immunoprecipitated with anti-FLAG antibodies and then analyzed by immunoblot using the indicated antibodies. (D) Quantitation of immunoprecipitated PP2A-Cα/β signal intensities from immunoblots in C and normalized to input PP2A-Cα/β protein (mean ± SD; three independent experiments). **P < 0.01. (E) HeLa cells were treated with 1 µM apilimod (or DMSO vehicle control) for 2 h. Cells were fixed and stained with the indicated antibodies and then analyzed by immunofluorescence microscopy. Bar, 10 µm. (F) Quantitation of colocalization ratio between mTOR and LAMP1 in E (mean ± SD; n > 40 cells from three independent experiments). **P < 0.01.