Fig. 7.

F-SLOH activates transcription factor EB (TFEB) promoting autophagy and lysosomal biogenesis in cellular models. (A&B) The ultrastructure of the autolysosome displayed in electron micrograph indicates that F-SLOH treatment in the HT-22 cells increased the lysosome number and autolysosome formation in comparison to the control and Torin1. (C) HT-22 (6.25, 12.5 and 25 μM) cells were treated with F-SLOH or Torin1 for 24 h increased the protein expression levels of LC3B-II using western blotting and its corresponding quantification. (D) The HT-22 cells were transfected with stably expressing tf-LC3 plasmids for 48 h and then treated with F-SLOH (25 μM) for 24 h and compared with Torin 1 and the lysosomal inhibitor chloroquine (CQ)(20 μM). The stained cells were pictured using confocal microscope. The autolysosome and autophagosome puncta in the HT-22 cells were quantified accordingly. (E) F-SLOH and Torin1 were treated in HT-22 cells for 24 h in the presence or absence of CQ. F-SLOH treatment increased the protein expression levels of LC3B-II and its corresponding quantification. (F) F-SLOH treatment in HT-22 cells for 24 h increased the protein expression levels of LAMP1 and mature Cathepsin D in a dose-dependent manner and its corresponding quantification. (G) F-SLOH or Torin1 treatment in HT-22 cells for 24 h and the cells were incubated with LysoTracker Green DND-99 (75 nm) for 1 h. The stained cells were detected using flow cytometer and its quantified data is shown in figure. (H) F-SLOH (25 μM) or Torin1 treatment in microglial cells for 24 h were stained for TFE3 protein expression and pictured in confocal microscope and its corresponding quantification. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)