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. 2020 Feb 22;21(4):1515. doi: 10.3390/ijms21041515

Figure 1.

Figure 1

Figure 1

Compound E4 promoted the nuclear translocation of endogenous and exogenous transcription factor EB (TFEB) in vitro. (A) Chemical structure of E4. (B) E4 significantly promoted the nuclear intensity of endogenous TFEB in N2a cells compared to control (0.1% DMSO). After being treated with E4 (1 μM) for 24 h, N2a cells were fixed and stained with the anti-TFEB antibody (red) and DAPI (blue). Representative images are shown. Scale bar: 15 μm. (C) E4 significantly promoted the nuclear intensity of TFEB in CF-7 cells compared to control (0.1% DMSO). After being treated with E4 (1 μM) and positive control Torin1 (250 nM) for 24 h, HeLa cells stably expressing 3XFlag-TFEB (CF-7) were fixed and stained with the anti-Flag antibody (green) and DAPI (blue). Representative images are shown. Scale bar: 15 μm. Quantification of the number of cells with nuclear TFEB localization. Data are presented as mean ± SEM of three replicates in a representative experiment. At least 100 cells were analyzed in each treatment group. (D) The expression of Flag-TFEB in the cytosol and the nucleus in CF-7 cells were detected by Western blotting after being treated with indicated compounds for 6 h. (E) Relative intensity of TFEB is normalized to that of GAPDH and H3. Data are presented as mean ± SEM of three replicates in a representative experiment. * p < 0.05, ** p < 0.01, *** p < 0.001.