Fig. 2: Impaired TFEB-regulated lysosomal dysregulation contributes to NPC senescence. | Cell Death & Differentiation

Fig. 2: Impaired TFEB-regulated lysosomal dysregulation contributes to NPC senescence.

From: Lysine methylation of PPP1CA by the methyltransferase SUV39H2 disrupts TFEB-dependent autophagy and promotes intervertebral disc degeneration

Fig. 2: Impaired TFEB-regulated lysosomal dysregulation contributes to NPC senescence.

a Representative images and quantification of total LysoTracker Red in NPCs with the indicated treatment (Scale bar: 10 μm). b Representative images and quantification of Magic Red dye in NPCs with the indicated treatment (Scale bar: 10 μm). c IF staining of TFEB in NPCs with the indicated treatment and the quantification of TFEB expression in the nucleus labeled with DAPI (Scale bar: 10 μm). d Cellular fractionation analysis of TFEB in NPCs with the indicated treatment. e Western blot analysis of p-TFEB and TFEB in human NPCs with the indicated treatment. f Western blot analysis of p53, p21, p16 in human NPCs transfected with si-TFEB. g, h Western blot analysis of LC3/p62 in human NPCs transfected with si-TFEB. i IF staining of TFEB in NPCs with the indicated treatment (Scale bar: 10 μm). j RT-qPCR analysis for TFEB target genes (TPP1, CTSF, MCOLN1, CTSD, LAMP1, ATG9A, WIPI1, ATG10, LC3B, SQSTM1) in NPCs with the indicated treatment. k Western blot analysis of p53, p21, p16 in human NPCs with the indicated treatment. l SA‐β‐gal activity staining and analysis of NPCs with the indicated treatment (Scale bar: 100 μm). m EdU incorporation assay and analysis of NPCs with the indicated treatment (Scale bar: 100 μm). Data are expressed as mean ± SD. *p < 0.05, **p < 0.01, **p < 0.001, ****p < 0.0001, ns not significant, two-tailed unpaired t-test and and one-way ANOVA.

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