Fig. 3
From: TFEB overexpression alleviates autophagy-lysosomal deficits caused by progranulin insufficiency
TFEB Overexpression in Wild-type Mouse Brain Increases Levels of Lysosomal Proteins. (a), We injected AAV2 vectors expressing either GFP or HA-tagged codon-optimized mouse TFEB into the ventroposteromedial/lateral nuclei of the thalamus (VPM/VPL) of wild-type mice. After 8 weeks, we collected brains for analysis of TFEB expression and lysosomal proteins. (b–d), AAV-TFEB primarily transduced neurons, based on immunostaining for the HA tag and markers of neurons (NeuN), astrocytes (S100β), and microglia (Iba1). (e), TFEB overexpression in thalamus was further confirmed by immunoblot (t test, p = 0.0373). (f), Immunoblot of thalamic tissue confirmed increases in lysosomal proteins in TFEB-treated mice (MANOVA effect of TFEB, p = 0.006), with increases in LAMP-1 (t test, p = 0.0141), mature cathepsin D (t test, p = 0.0017), and immature cathepsin D (t test, p = 0.0003). (g), However, analysis of the LC3-II/LC3-I ratio (t test, p = 0.6163) and p62 (t test, p = 0.5056) did not reveal signs of increased autophagy. n = 6–7 mice per group. Scale bars in (b–d) represent 25 μm. Panel (a) created using Biorender.com. (Arrant, A. (2025) https://BioRender.com/flnev93 and https://BioRender.com/6cia20y). Full images of immunoblots are shown in Fig. S4.
