Skip to main content
. 2021 Apr 1;11(10):3134–3149. doi: 10.1016/j.apsb.2021.03.039

Figure 5.

Figure 5

TBM-1 triggers TFEB nuclear translocation. (A) H157 cells were transfected with TFEB siRNA, TFE3 siRNA, MITF siRNA, followed by TBM-1 (5 μmol/L) treatment for 12 h. Images and quantification of lysosomes (indicated as LysoTracker staining) were determined (scale bar, 50 μm). (B) Images and quantification of the subcellular locations of EGFP-TFEB, EGFP-MITF in H157 cells treatment with DMSO or TBM-1 (5 μmol/L) for 12 h. (scale bar, 20 μm). (C) Immunoblotting detecting the cytosolic and nuclear distribution of TFEB in H157and A375 cells treatment with DMSO or TBM-1 (5 μmol/L, 12 h). Quantifications of nuclear TFEB to H3 were shown at the right. (D) Immunoblotting determining the levels of TFEB Ser211 phosphorylation in H157 and A375 cells treated with indicated TBM-1 for 12 h. Quantifications of p-TFEB to TFEB are shown at the right. (E) H157 and A375 cells were treated with 5 μmol/L TBM-1 for 12 h. 14-3-3 levels were measured by immunoblotting after immunoprecipitation of TFEB from cell lysates. Quantifications of 14-3-3 to TFEB were shown. (F) and (G) Control siRNA or TFEB siRNA were transfected to H157 cells for 2 days, followed by DMSO or TBM-1 (5 μmol/L) treatment for 12 h. The levels of LAMP1, PD-L1 (F) and relative lysosomal NAG activity (G) were determined. Data are presented as mean ± SD, n = 3; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001, ns, no significant.