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<t>A.</t> <t>Lysosomal</t> acidification was assessed by measuring the uptake of the fluorescent dye pHLys Green in cells treated with the TFEB activator curcumin analog <t>C1.</t> The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p** < 0.01, ns; not significant. B . Similarly, lysosomal abundance was measured using LysoPrime Deep Red. The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p* < 0.05, p** < 0.01. C . Cells expressing TauRD-GFP were pretreated with the TFEB activator curcumin analog C1 for 24 h and then seeded with PFFs. Cells were subsequently treated with curcumin analog C1 for 48 h, followed by assessment of TauRD-GFP aggregation. D . Results of the TauRD-GFP aggregation assay in WT and G3BP1 KO cells are shown. Open bars (DMSO) represent DMSO-treated cells, and closed bars (C1) represent cells treated with curcumin analog C1. The data are expressed as means ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-hoc test. p*** < 0.001, ns; not significant.
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<t>A.</t> <t>Lysosomal</t> acidification was assessed by measuring the uptake of the fluorescent dye pHLys Green in cells treated with the TFEB activator curcumin analog <t>C1.</t> The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p** < 0.01, ns; not significant. B . Similarly, lysosomal abundance was measured using LysoPrime Deep Red. The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p* < 0.05, p** < 0.01. C . Cells expressing TauRD-GFP were pretreated with the TFEB activator curcumin analog C1 for 24 h and then seeded with PFFs. Cells were subsequently treated with curcumin analog C1 for 48 h, followed by assessment of TauRD-GFP aggregation. D . Results of the TauRD-GFP aggregation assay in WT and G3BP1 KO cells are shown. Open bars (DMSO) represent DMSO-treated cells, and closed bars (C1) represent cells treated with curcumin analog C1. The data are expressed as means ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-hoc test. p*** < 0.001, ns; not significant.
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A. Lysosomal acidification was assessed by measuring the uptake of the fluorescent dye pHLys Green in cells treated with the TFEB activator curcumin analog C1. The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p** < 0.01, ns; not significant. B . Similarly, lysosomal abundance was measured using LysoPrime Deep Red. The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p* < 0.05, p** < 0.01. C . Cells expressing TauRD-GFP were pretreated with the TFEB activator curcumin analog C1 for 24 h and then seeded with PFFs. Cells were subsequently treated with curcumin analog C1 for 48 h, followed by assessment of TauRD-GFP aggregation. D . Results of the TauRD-GFP aggregation assay in WT and G3BP1 KO cells are shown. Open bars (DMSO) represent DMSO-treated cells, and closed bars (C1) represent cells treated with curcumin analog C1. The data are expressed as means ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-hoc test. p*** < 0.001, ns; not significant.

Journal: bioRxiv

Article Title: G3BP1 Maintains Lysosomal Homeostasis to Limit Tau Aggregate Accumulation

doi: 10.1101/2025.08.29.672259

Figure Lengend Snippet: A. Lysosomal acidification was assessed by measuring the uptake of the fluorescent dye pHLys Green in cells treated with the TFEB activator curcumin analog C1. The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p** < 0.01, ns; not significant. B . Similarly, lysosomal abundance was measured using LysoPrime Deep Red. The data are presented as means ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. p* < 0.05, p** < 0.01. C . Cells expressing TauRD-GFP were pretreated with the TFEB activator curcumin analog C1 for 24 h and then seeded with PFFs. Cells were subsequently treated with curcumin analog C1 for 48 h, followed by assessment of TauRD-GFP aggregation. D . Results of the TauRD-GFP aggregation assay in WT and G3BP1 KO cells are shown. Open bars (DMSO) represent DMSO-treated cells, and closed bars (C1) represent cells treated with curcumin analog C1. The data are expressed as means ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-hoc test. p*** < 0.001, ns; not significant.

Article Snippet: Lysosomal biogenesis was promoted using curcumin analog C1 (TFEB activator 1; MedChemExpress, Monmouth Junction, NJ).

Techniques: Expressing

A schematic model illustrating the differences in lysosomal homeostasis between WT and G3BP1 KO cells. In WT cells (upper panels), G3BP1 cooperates with ESCRT and other factors to repair damaged lysosomes and enhance lysosomal activity, thereby promoting the degradation of Tau aggregates and PFFs. In contrast, in G3BP1 KO cells, these functions are impaired, leading to reduced lysosomal activity and the accumulation of Tau aggregates. However, activation of TFEB by the curcumin analog C1 promotes lysosomal biogenesis and suppresses the accumulation of Tau aggregates.

Journal: bioRxiv

Article Title: G3BP1 Maintains Lysosomal Homeostasis to Limit Tau Aggregate Accumulation

doi: 10.1101/2025.08.29.672259

Figure Lengend Snippet: A schematic model illustrating the differences in lysosomal homeostasis between WT and G3BP1 KO cells. In WT cells (upper panels), G3BP1 cooperates with ESCRT and other factors to repair damaged lysosomes and enhance lysosomal activity, thereby promoting the degradation of Tau aggregates and PFFs. In contrast, in G3BP1 KO cells, these functions are impaired, leading to reduced lysosomal activity and the accumulation of Tau aggregates. However, activation of TFEB by the curcumin analog C1 promotes lysosomal biogenesis and suppresses the accumulation of Tau aggregates.

Article Snippet: Lysosomal biogenesis was promoted using curcumin analog C1 (TFEB activator 1; MedChemExpress, Monmouth Junction, NJ).

Techniques: Activity Assay, Activation Assay