Fig. 4
From: TFEB overexpression alleviates autophagy-lysosomal deficits caused by progranulin insufficiency
TFEB Overexpression Reduces Lysosomal Storage Material in Grn–/– Mice. (a), AAV-TFEB expressed HA-tagged TFEB throughout the ventroposteromedial/lateral thalamic nuclei of Grn–/– mice. (b–d), Similar to wild-type mice, AAV-TFEB primarily transduced neurons and increased TFEB levels in the thalamus (e, ANOVA effect of TFEB, p < 0.0001, *** = p = 0.0001 and **** = p < 0.0001 by Holm-Sidak post-hoc test). (f), In contrast to wild-type mice, AAV-TFEB did not increase LAMP-1 or cathepsin D (CatD) in Grn–/– mice (MANOVA effect of TFEB, p = 0.467). AAV-TFEB also did not alter the LC3-II/LC3-I ratio (g, ANOVA effect of TFEB, p = 0.2413) or p62 levels (g, ANOVA effect of TFEB, p = 0.9191) in Grn–/– mice. For (e–g), n = 10–15 mice/group. i, However, AAV-TFEB did increase levels of multiple lysosomal transcripts as measured by qPCR (MANOVA effect of TFEB, p = 0.04, * = p < 0.05, ** = p < 0.01 by t test, n = 6–10 samples/group). (j), AAV-TFEB reduced immunoreactivity for SCMAS, a marker of lysosomal storage material that is elevated in Grn–/– thalamus (ANOVA effect of TFEB, p = 0.0063, * = p = 0.0039 by Holm-Sidak post-hoc test, n = 13–27 mice/group). (k), AAV-TFEB produced a similar trend for autofluorescent lipofuscin (ANOVA effect of TFEB, p = 0.2730), which exhibited greater variability, especially in the uninjected group. Representative 10X images of SCMAS immunostaining and autofluorescence are shown in l with 100 μm scale bars. SCMAS = subunit C of mitochondrial ATP synthase. Full images of immunoblots are shown in Fig. S5.
