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. 2015 Feb 18;35(6):956–976. doi: 10.1128/MCB.01091-14

Fig 6.

Fig 6

Exogenous beclin-1 inactivates TFEB. (A and B) Representative flow cytometric tracings (A) and quantitation (B) of LysoTracker fluorescence in NRCMs transduced with beclin-1 or LacZ (MOI = 100; 48 h) (n = 6). (C) Immunoblot (IB) demonstrating subcellular distribution of TFEB in NRCMs transduced with HA-TFEB (MOI = 1), treated as described for panel A, and subjected to subcellular fractionation into cytoplasmic and nuclear fractions. The data are representative of 2 experiments. (D and E) Immunoblot (D) and quantitation (E) of TFEB abundance in NRCMs transduced with HA-TFEB (MOI = 1) and treated as described for panel A, with or without MG-132 (10 μmol/liter; 24 h) (n = 4). (F) Immunoblots for phosphorylation of S6K, 4EBP-1, and mTOR in NRCMs transduced with beclin-1 or LacZ (MOI = 100; 48 h), with or without torin-1 (250 nmol/liter; 1 h). (G) Representative immunoblot (n = 2) depicting coimmunoprecipitation (IP) of endogenous TSC1 (left) and beclin-1 (right) in MEFs expressing beclin-1–HA and HA-TSC1, respectively. (H to J) Immunoblot (H) and quantitation of TSC1 (I) and TSC2 (J) in NRCMs treated as described for panel A (n = 4).