frap module Search Results


zen black module frap efficiency analysis  (Carl Zeiss)
93
Carl Zeiss zen black module frap efficiency analysis
Zen Black Module Frap Efficiency Analysis, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/pmc12876695-100-0-7?v=Carl+Zeiss
Average 93 stars, based on 1 article reviews
zen black module frap efficiency analysis - by Bioz Stars, 2026-08
93/100 stars
  Buy from Supplier
frap module  (Carl Zeiss)
94
Carl Zeiss frap module
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
Frap Module, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/bio_rxiv__2023__05__02__538687-192-6-10?v=Carl+Zeiss
Average 94 stars, based on 1 article reviews
frap module - by Bioz Stars, 2026-08
94/100 stars
  Buy from Supplier
2d-visifrap galvo frap module  (VISITRON Inc)
90
VISITRON Inc 2d-visifrap galvo frap module
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
2d Visifrap Galvo Frap Module, supplied by VISITRON Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/pmc09020716-264-50-71?v=VISITRON+Inc
Average 90 stars, based on 1 article reviews
2d-visifrap galvo frap module - by Bioz Stars, 2026-08
90/100 stars
  Buy from Supplier
frap module  (Rapp OptoElectronic GmbH)
90
Rapp OptoElectronic GmbH frap module
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
Frap Module, supplied by Rapp OptoElectronic GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/pm40632866-235-5-5?v=Rapp+OptoElectronic+GmbH
Average 90 stars, based on 1 article reviews
frap module - by Bioz Stars, 2026-08
90/100 stars
  Buy from Supplier
frap module within the visitron software  (VISITRON Inc)
90
VISITRON Inc frap module within the visitron software
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
Frap Module Within The Visitron Software, supplied by VISITRON Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/pm40164947-620-10-14?v=VISITRON+Inc
Average 90 stars, based on 1 article reviews
frap module within the visitron software - by Bioz Stars, 2026-08
90/100 stars
  Buy from Supplier
multi-point frap/photo-activation module  (VISITRON Inc)
90
VISITRON Inc multi-point frap/photo-activation module
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
Multi Point Frap/Photo Activation Module, supplied by VISITRON Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/pmc06705222-343-47-51?v=VISITRON+Inc
Average 90 stars, based on 1 article reviews
multi-point frap/photo-activation module - by Bioz Stars, 2026-08
90/100 stars
  Buy from Supplier
515nm laser metamorph with frap module  (MetaMorph Inc)
90
MetaMorph Inc 515nm laser metamorph with frap module
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
515nm Laser Metamorph With Frap Module, supplied by MetaMorph Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/pm38604167-452-12-13?v=MetaMorph+Inc
Average 90 stars, based on 1 article reviews
515nm laser metamorph with frap module - by Bioz Stars, 2026-08
90/100 stars
  Buy from Supplier
ilas2 scanner system  (Gataca Inc)
86
Gataca Inc ilas2 scanner system
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
Ilas2 Scanner System, supplied by Gataca Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/bio_rxiv__64898__2026__03__23__713115-205-5-8?v=Gataca+Inc
Average 86 stars, based on 1 article reviews
ilas2 scanner system - by Bioz Stars, 2026-08
86/100 stars
  Buy from Supplier
ilas frap illumination module  (Roper Scientific Inc)
86
Roper Scientific Inc ilas frap illumination module
GATA3 undergoes LLPS in cells and in vitro. ( A ) <t>Representative</t> <t>confocal</t> microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in <t>FRAP</t> assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.
Ilas Frap Illumination Module, supplied by Roper Scientific Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/frap+module/bio_rxiv__126151-272-45-49?v=Roper+Scientific+Inc
Average 86 stars, based on 1 article reviews
ilas frap illumination module - by Bioz Stars, 2026-08
86/100 stars
  Buy from Supplier

Image Search Results


GATA3 undergoes LLPS in cells and in vitro. ( A ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in FRAP assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.

Journal: bioRxiv

Article Title: A Zinc finger (ZnF) transcription factor exerts its multifaceted function through ZnF-regulated phase separation

doi: 10.1101/2023.05.02.538687

Figure Lengend Snippet: GATA3 undergoes LLPS in cells and in vitro. ( A ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged wild type GATA3 (GATA3 WT ). Scale bar, 5 μm. ( B ) Representative confocal microscopy images of Hela cells with ectopic overexpression of mEGFP-tagged GATA3 WT . Scale bar, 2 μm. ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 WT after treatment with 10% 1,6-hexanediol (1.6-Hex) compared to control. Scale bar,2 μm. ( D ) Representative images of mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in FRAP assay. Scale bar, 2 μm. ( E ) Normalized fluorescence recovery of mEGFP-tagged GATA3 WT droplets in HEK293T cells as shown in ( D ), t 1/2 , means half-life of recovery time. ( F ) Representative images of 10 μM purified MBP-GATA3 WT protein without mEGFP tag in tubes treated with 3C protease treatment compared to control. ( G ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein without mEGFP tag with 3C protease treatment compared to control. Scale bar, 50 μm. ( H ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag compared to 10 μM purified MBP-mEGFP protein post the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( I ) FRAP assay for droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Right, representative images of droplets across time in the FRAP assay. Light, normalized fluorescence recovery of droplets in FRAP assay. Scale bar, 1μm. ( J ) Representative images of droplet fusion events using 10 μM purified MBP-GATA3 WT protein with mEGFP tag after its MBP was cleaved by 3C protease treatment. Scale bar, 5 μm.

Article Snippet: FRAP assay was conducted using the FRAP module of the Zeiss LSM710/810 ZEN blue confocal microscopy system.

Techniques: In Vitro, Confocal Microscopy, Over Expression, FRAP Assay, Fluorescence, Purification

ZnF2 and its amino acids R329 and R330 regulate the LLPS of GATA3. ( A ) A representative image elucidating the complex structure of ZnF1 in GATA3 binding to DNA (PDB: 4HC9). ( B ) A representative image elucidating the complex structure of ZnF2 in GATA3 binding to DNA (PDB: 4HC7). ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 mutants compared to mEGFP-tagged GATA3 WT as indicated in the figure. Scale bar, 2 μm. ( D ) Quantification of the number of droplets per cell shown in ( C ). Data are mean□±□s.d.. ***, p<0.001; ns, not significant (t-test). ( E ) Representative images of droplets formed by 10 μM purified MBP-ZnF2 2R-A protein with mEGFP tag after the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( F ) Representative images of mEGFP-tagged ZnF2 2R-A droplets compared to mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in FRAP assay. Scale bars, 2 μm. ( G ) Normalized fluorescence recovery of mEGFP-tagged ZnF2 2R-A (Red) droplets compared to mEGFP-tagged GATA3 WT (Blue) droplets in HEK293T cells as shown in ( D ), t 1/2 , half-life of recovery time. ( H ) Quantification the size of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag and the MBP cleaved by 3C protease treatment in the visual fields, and DNA treatment was indicated in the figure. Blue, noGATA-DNA added. Red, GATA-DNA added. Black, no DNA added. GATA3-mEGFP protein concentration, 10 μM. ( I ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag and the MBP cleaved by 3C protease treatment with 20 μM noGATA-DNA or 20 μM GATA-DNA added compared to no DNA added. ( J ) Representative images of droplets formed by 10 μM purified mEGFP-tagged ZnF2 2R-A protein compared to 10 μM purified mEGFP-tagged GATA3 WT protein with 20 μM noGATA-DNA or 20 μM GATA-DNA added as indicated in the figure across time in FRAP assay. The MBP of purified proteins was cleaved by 3C protease treatment. Scale bars, 10 μm. ( K ) Normalized fluorescence recovery of droplets as shown in ( D ). Blue, 20 μM no-GATA DNA added. Red, 20 μM GATA DNA added. Black, no DNA added.

Journal: bioRxiv

Article Title: A Zinc finger (ZnF) transcription factor exerts its multifaceted function through ZnF-regulated phase separation

doi: 10.1101/2023.05.02.538687

Figure Lengend Snippet: ZnF2 and its amino acids R329 and R330 regulate the LLPS of GATA3. ( A ) A representative image elucidating the complex structure of ZnF1 in GATA3 binding to DNA (PDB: 4HC9). ( B ) A representative image elucidating the complex structure of ZnF2 in GATA3 binding to DNA (PDB: 4HC7). ( C ) Representative confocal microscopy images of HEK293T cells with ectopic overexpression of mEGFP-tagged GATA3 mutants compared to mEGFP-tagged GATA3 WT as indicated in the figure. Scale bar, 2 μm. ( D ) Quantification of the number of droplets per cell shown in ( C ). Data are mean□±□s.d.. ***, p<0.001; ns, not significant (t-test). ( E ) Representative images of droplets formed by 10 μM purified MBP-ZnF2 2R-A protein with mEGFP tag after the MBP cleaved by 3C protease treatment. Scale bar, 10 μm. ( F ) Representative images of mEGFP-tagged ZnF2 2R-A droplets compared to mEGFP-tagged GATA3 WT droplets in HEK293T cells across time in FRAP assay. Scale bars, 2 μm. ( G ) Normalized fluorescence recovery of mEGFP-tagged ZnF2 2R-A (Red) droplets compared to mEGFP-tagged GATA3 WT (Blue) droplets in HEK293T cells as shown in ( D ), t 1/2 , half-life of recovery time. ( H ) Quantification the size of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag and the MBP cleaved by 3C protease treatment in the visual fields, and DNA treatment was indicated in the figure. Blue, noGATA-DNA added. Red, GATA-DNA added. Black, no DNA added. GATA3-mEGFP protein concentration, 10 μM. ( I ) Representative images of droplets formed by 10 μM purified MBP-GATA3 WT protein with mEGFP tag and the MBP cleaved by 3C protease treatment with 20 μM noGATA-DNA or 20 μM GATA-DNA added compared to no DNA added. ( J ) Representative images of droplets formed by 10 μM purified mEGFP-tagged ZnF2 2R-A protein compared to 10 μM purified mEGFP-tagged GATA3 WT protein with 20 μM noGATA-DNA or 20 μM GATA-DNA added as indicated in the figure across time in FRAP assay. The MBP of purified proteins was cleaved by 3C protease treatment. Scale bars, 10 μm. ( K ) Normalized fluorescence recovery of droplets as shown in ( D ). Blue, 20 μM no-GATA DNA added. Red, 20 μM GATA DNA added. Black, no DNA added.

Article Snippet: FRAP assay was conducted using the FRAP module of the Zeiss LSM710/810 ZEN blue confocal microscopy system.

Techniques: Binding Assay, Confocal Microscopy, Over Expression, Purification, FRAP Assay, Fluorescence, Protein Concentration