tfeb (Bethyl)
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Structured Review
Bethyl
tfeb
![( A ) Volcano plots of proteins from whole-cell proteomics in <t>TFEB-3xFlag</t> versus wild-type (WT) U2OS cells. Significantly altered proteins: dark [FDR < 0.05, log 2 fold change (FC) > |1|] and light (FDR < 0.05, 0 < |log 2 FC| < |1|) red/blue (FDR-corrected two-sided t test, N = 4). E3 ligases are highlighted in black (table S1). ( B ) Western blot of indicated proteins in control and TFEB-3xFlag U2OS cells ( N = 3) with or without BafA1 (200 nM, 4 hours). Quantification normalized <t>to</t> <t>β-actin;</t> means ± SEM. One-way ANOVA: P = 0.0003. Sidak’s test: ** P < 0.005. M r , relative molecular mass. ( C to E ) Coimmunofluorescence of SQSTM1 (red) with (C) NBR1, (D) TAX1BP1, or (E) LC3B (green) in CTRL and TFEB-GFP (purple) cells. Scale bars, 10 μm (insets, 2 μm). Quantification of puncta per cell; means ± SEM ( N = 3, n = 24 to 30 cells). Student’s unpaired t test: ** P < 0.005 for NBR1, * P < 0.05 for TAX1BP1 and LC3B. ( F ) Electron micrograph of TFEB-GFP U2OS cell labeled for SQSTM1 (nanogold). ( G ) CLEM of TFEB-GFP U2OS cells starved in HBSS (2 hours) and labeled for SQSTM1 (red) and LC3B (green). The arrow indicates membranes surrounding an SQSTM1- and LC3B-positive structure. ( H ) Coimmunofluorescence of SQSTM1 (red) and LC3B (green) in FLCN KO HeLa cells ± siSQSTM1 (100 nM for 48 hours). Scale bars, 10 μm (insets, 5 μm). Quantification of LC3B puncta per cell; means ± SEM ( N = 3, n = 30 cells). Student’s unpaired t test: * P < 0.05.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7066/pmc12757066/pmc12757066__sciadv.aea9302-f1.jpg)
Tfeb, supplied by Bethyl, used in various techniques. Bioz Stars score: 96/100, based on 361 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/tfeb/product/Bethyl
Average 96 stars, based on 361 article reviews
Tfeb, supplied by Bethyl, used in various techniques. Bioz Stars score: 96/100, based on 361 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/tfeb/product/Bethyl
Average 96 stars, based on 361 article reviews
tfeb - by Bioz Stars,
2026-04
96/100 stars
Images
1) Product Images from "TFEB coordinates autophagosome biogenesis and ribophagy during starvation via SQSTM1"
Article Title: TFEB coordinates autophagosome biogenesis and ribophagy during starvation via SQSTM1
Journal: Science Advances
doi: 10.1126/sciadv.aea9302
Figure Legend Snippet: ( A ) Volcano plots of proteins from whole-cell proteomics in TFEB-3xFlag versus wild-type (WT) U2OS cells. Significantly altered proteins: dark [FDR < 0.05, log 2 fold change (FC) > |1|] and light (FDR < 0.05, 0 < |log 2 FC| < |1|) red/blue (FDR-corrected two-sided t test, N = 4). E3 ligases are highlighted in black (table S1). ( B ) Western blot of indicated proteins in control and TFEB-3xFlag U2OS cells ( N = 3) with or without BafA1 (200 nM, 4 hours). Quantification normalized to β-actin; means ± SEM. One-way ANOVA: P = 0.0003. Sidak’s test: ** P < 0.005. M r , relative molecular mass. ( C to E ) Coimmunofluorescence of SQSTM1 (red) with (C) NBR1, (D) TAX1BP1, or (E) LC3B (green) in CTRL and TFEB-GFP (purple) cells. Scale bars, 10 μm (insets, 2 μm). Quantification of puncta per cell; means ± SEM ( N = 3, n = 24 to 30 cells). Student’s unpaired t test: ** P < 0.005 for NBR1, * P < 0.05 for TAX1BP1 and LC3B. ( F ) Electron micrograph of TFEB-GFP U2OS cell labeled for SQSTM1 (nanogold). ( G ) CLEM of TFEB-GFP U2OS cells starved in HBSS (2 hours) and labeled for SQSTM1 (red) and LC3B (green). The arrow indicates membranes surrounding an SQSTM1- and LC3B-positive structure. ( H ) Coimmunofluorescence of SQSTM1 (red) and LC3B (green) in FLCN KO HeLa cells ± siSQSTM1 (100 nM for 48 hours). Scale bars, 10 μm (insets, 5 μm). Quantification of LC3B puncta per cell; means ± SEM ( N = 3, n = 30 cells). Student’s unpaired t test: * P < 0.05.
Techniques Used: Western Blot, Control, Labeling
![( A ) Schematic representation of the SQSTM1 promoter with putative TFEB binding sites. Regions 1 and 2 were cloned into the pGL3-basic luciferase reporter plasmid, and the luciferase activity was determined. Means ± SEM of N = 3. One-way ANOVA: P = 0.0016. Sidak’s test: ** P < 0.005. ns, not significant. TSS, transcription start site; 5′UTR, 5′ untranslated region; A.U., arbitrary units. ( B ) qRT-PCR analysis of SQSTM1 expression in CTRL and ΔCLEAR HeLa cells with or without TFEB overexpression (TFEB OE). Fold change normalized to HPRT and expressed relative to CTRL. Means ± SEM of N = 3. One-way ANOVA, P < 0.0001. Sidak’s test: *** P < 0.0001. ( C ) Immunofluorescence of SQSTM1 (red) and TFEB-GFP (purple) in WT and ΔCLEAR HeLa cells overexpressing TFEB-GFP. Nuclei stained with DAPI (blue). Scale bars, 10 μm (insets, 2 μm). Quantification of SQSTM1 puncta per cell; means ± SEM ( N = 3, n = 40 cells). Student’s unpaired t test: * P < 0.05. ( D ) Western blot analysis of indicated proteins in WT and ΔCLEAR HeLa cells infected with TFEB3xFlag, with or without BafA1 (200 nM, 4 hours). Quantification of LC3BII normalized to β-actin; means ± SEM ( N = 4). One-way ANOVA: *** P < 0.0001. Sidak’s test: *** P < 0.0005; ** P < 0.005; * P < 0.05. ( E ) Coimmunofluorescence staining of LC3B (green), SQSTM1 (red), and TFEB-GFP (purple) in WT and ΔCLEAR HeLa cells overexpressing TFEB-GFP. Scale bars, 10 μm. Quantification of LC3B puncta per cell; means ± SEM [ N = 3, n = 41 (HeLa) and n = 40 (ΔCLEAR HeLa) cells]. Student’s unpaired t test: *** P = 0.001.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7066/pmc12757066/pmc12757066__sciadv.aea9302-f2.jpg "... Schematic representation of the SQSTM1 promoter with putative TFEB binding sites. Regions 1 and 2 were cloned ...")
Figure Legend Snippet: ( A ) Schematic representation of the SQSTM1 promoter with putative TFEB binding sites. Regions 1 and 2 were cloned into the pGL3-basic luciferase reporter plasmid, and the luciferase activity was determined. Means ± SEM of N = 3. One-way ANOVA: P = 0.0016. Sidak’s test: ** P < 0.005. ns, not significant. TSS, transcription start site; 5′UTR, 5′ untranslated region; A.U., arbitrary units. ( B ) qRT-PCR analysis of SQSTM1 expression in CTRL and ΔCLEAR HeLa cells with or without TFEB overexpression (TFEB OE). Fold change normalized to HPRT and expressed relative to CTRL. Means ± SEM of N = 3. One-way ANOVA, P < 0.0001. Sidak’s test: *** P < 0.0001. ( C ) Immunofluorescence of SQSTM1 (red) and TFEB-GFP (purple) in WT and ΔCLEAR HeLa cells overexpressing TFEB-GFP. Nuclei stained with DAPI (blue). Scale bars, 10 μm (insets, 2 μm). Quantification of SQSTM1 puncta per cell; means ± SEM ( N = 3, n = 40 cells). Student’s unpaired t test: * P < 0.05. ( D ) Western blot analysis of indicated proteins in WT and ΔCLEAR HeLa cells infected with TFEB3xFlag, with or without BafA1 (200 nM, 4 hours). Quantification of LC3BII normalized to β-actin; means ± SEM ( N = 4). One-way ANOVA: *** P < 0.0001. Sidak’s test: *** P < 0.0005; ** P < 0.005; * P < 0.05. ( E ) Coimmunofluorescence staining of LC3B (green), SQSTM1 (red), and TFEB-GFP (purple) in WT and ΔCLEAR HeLa cells overexpressing TFEB-GFP. Scale bars, 10 μm. Quantification of LC3B puncta per cell; means ± SEM [ N = 3, n = 41 (HeLa) and n = 40 (ΔCLEAR HeLa) cells]. Student’s unpaired t test: *** P = 0.001.
Techniques Used: Binding Assay, Clone Assay, Luciferase, Plasmid Preparation, Activity Assay, Quantitative RT-PCR, Expressing, Over Expression, Immunofluorescence, Staining, Western Blot, Infection
![( A ) The UBQ-HA interactome in TFEB-3xFlag U2OS cells identified ribosomal proteins as a prominent category (table S2). ( B ) Volcano plot of ubiquitinated peptides by diGly proteomics in TFEB-3xFlag versus WT U2OS; significant changes in red/blue (FDR < 0.05, log 2 FC > 1 or < 1); two-sided t test, N = 4 (table S1). ( C ) Top five significant GO CC terms among 465 up-regulated proteins; enrichment score (ES) shown (FDR = 10% and ES > 1.5; table S5). ( D ) Heatmap of TFEB-regulated E3 ligase (FDR-corrected t test, N = 4; table S1). ( E ) ZNF598 promoter schematic showing putative TFEB binding sites. ( F and G ) qRT-PCR of ZNF598 in mock, TFEB-GFP, or siTFEB-TFE3 U2OS ± HBSS (4 hours); normalized fold change (means ± SEM, N = 3 or 4; ** P < 0.005; ANOVA: * P = 0.031; Sidak’s test: * P < 0.05). ( H ) Western blot of ZNF598 under indicated conditions; quantified versus β-actin (means ± SEM, N = 3), ** P < 0.005. ( I ) Heatmap of significantly HA-ubiquitinated ribosomal proteins (S0 = 0.1, FDR < 0.05, N = 4) under indicated conditions (red: up-regulated; blue: down-regulated; table S3). ( J ) Coimmunofluorescence of SQSTM1 (red) and RPS3 (green) in TFEB-GFP ± siZNF598 ; scale bars, 10 μm (insets, 2 μm). Quantification: SQSTM1-RPS3 colocalization (%) and SQSTM1 puncta per cell (means ± SEM of N = 3, n = 38), * P < 0.05. ( K ) Fluorescence microscopy of the RPS3 reporter in starved (ON) TFEB-3xFlag ± siZNF598 . Scale bars, 10 μm (insets, 2 μm). RFP intensity relative to scramble (means ± SEM of N = 3, n = 30), * P = 0.007. ( L ) FACS of the RPS3 WT or K214R reporter. Red fluorescence shift, means ± SEM [ N = 4 (WT), N = 6 (K214R)]. ANOVA: P = 0.0006 (WT), P = 0.01 (K214R). Sidak’s test: * P < 0.05; ** P < 0.005.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7066/pmc12757066/pmc12757066__sciadv.aea9302-f5.jpg "( A ) The UBQ-HA interactome in TFEB-3xFlag U2OS cells identified ribosomal proteins as a prominent ...")
Figure Legend Snippet: ( A ) The UBQ-HA interactome in TFEB-3xFlag U2OS cells identified ribosomal proteins as a prominent category (table S2). ( B ) Volcano plot of ubiquitinated peptides by diGly proteomics in TFEB-3xFlag versus WT U2OS; significant changes in red/blue (FDR < 0.05, log 2 FC > 1 or < 1); two-sided t test, N = 4 (table S1). ( C ) Top five significant GO CC terms among 465 up-regulated proteins; enrichment score (ES) shown (FDR = 10% and ES > 1.5; table S5). ( D ) Heatmap of TFEB-regulated E3 ligase (FDR-corrected t test, N = 4; table S1). ( E ) ZNF598 promoter schematic showing putative TFEB binding sites. ( F and G ) qRT-PCR of ZNF598 in mock, TFEB-GFP, or siTFEB-TFE3 U2OS ± HBSS (4 hours); normalized fold change (means ± SEM, N = 3 or 4; ** P < 0.005; ANOVA: * P = 0.031; Sidak’s test: * P < 0.05). ( H ) Western blot of ZNF598 under indicated conditions; quantified versus β-actin (means ± SEM, N = 3), ** P < 0.005. ( I ) Heatmap of significantly HA-ubiquitinated ribosomal proteins (S0 = 0.1, FDR < 0.05, N = 4) under indicated conditions (red: up-regulated; blue: down-regulated; table S3). ( J ) Coimmunofluorescence of SQSTM1 (red) and RPS3 (green) in TFEB-GFP ± siZNF598 ; scale bars, 10 μm (insets, 2 μm). Quantification: SQSTM1-RPS3 colocalization (%) and SQSTM1 puncta per cell (means ± SEM of N = 3, n = 38), * P < 0.05. ( K ) Fluorescence microscopy of the RPS3 reporter in starved (ON) TFEB-3xFlag ± siZNF598 . Scale bars, 10 μm (insets, 2 μm). RFP intensity relative to scramble (means ± SEM of N = 3, n = 30), * P = 0.007. ( L ) FACS of the RPS3 WT or K214R reporter. Red fluorescence shift, means ± SEM [ N = 4 (WT), N = 6 (K214R)]. ANOVA: P = 0.0006 (WT), P = 0.01 (K214R). Sidak’s test: * P < 0.05; ** P < 0.005.
Techniques Used: Ubiquitin Proteomics, Binding Assay, Quantitative RT-PCR, Western Blot, Fluorescence, Microscopy