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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used <t>for</t> <t>real-time</t> <t>PCR.</t> Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).
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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used <t>for</t> <t>real-time</t> <t>PCR.</t> Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).
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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used <t>for</t> <t>real-time</t> <t>PCR.</t> Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).
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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used <t>for</t> <t>real-time</t> <t>PCR.</t> Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).
Real Time Fluorescent Quantitative Pcr Kit, supplied by Vazyme Biotech Co, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used <t>for</t> <t>real-time</t> <t>PCR.</t> Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).
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quantitative real time pcr  (Roche)
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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used <t>for</t> <t>real-time</t> <t>PCR.</t> Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).
Quantitative Real Time Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used <t>for</t> <t>real-time</t> <t>PCR.</t> Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).
Quantitative Real Time Pcr Employed Chamq Universal Sybr Qpcr Master Mix, supplied by Vazyme Biotech Co, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used for real-time PCR. Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).

Journal: Molecules and Cells

Article Title: Tankyrase-1-mediated PARsylation directs TFEB partner switching to regulate selective Wnt target gene expression

doi: 10.1016/j.mocell.2026.100313

Figure Lengend Snippet: PARsylation of TFEB is crucial for its interaction with TCF-1/LEF-1 and Wnt-TFEB target gene expression. (A) TFEB with mutations at residues 237K and 274K is not PARsylated by TNKS1. (B, C) TFEB with mutations in the PARsylation sites does not bind to HA-TCF-1 (B) or HA-LEF-1 (C) in HEK293T cells. (D) The self-interaction of the TFEB mutant, which has mutations at PARsylation sites, was not reduced by the treatment of Wnt3a-CM. HEK293A cells were transfected with TFEB mutants carrying different tags and treated with Wnt3a-CM for 2 hours. (E) Treatment of HEK293A cells with Wnt3a-CM for 2 hours does not reduce the level of oligomer of TFEB PARsylation-defective mutants. (F) PARsylation of TFEB is necessary for Wnt-TFEB target gene expression. HEK293A cells were knocked down and transfected with the indicated constructs, then treated with Wnt3a-CM for 8 hours. The cells were then used for real-time PCR. Statistical analysis was performed using a 2-tailed unpaired t-test, showing exact P- values in each figure. Error bars indicate the standard deviation of triplicate measurements (technical replicates).

Article Snippet: Quantitative real-time PCR (qPCR) was conducted using THUNDERBIRD SYBR qPCR Mix (Toyobo, QPS-201) in accordance with the manufacturer’s instructions.

Techniques: Targeted Gene Expression, Mutagenesis, Transfection, Construct, Real-time Polymerase Chain Reaction, Standard Deviation