Journal: Cell reports
Article Title: GATA3 differentially regulates the transcriptome via zinc finger 2-modulated phase separation.
doi: 10.1016/j.celrep.2025.115702
Figure Lengend Snippet: Figure 1. GATA3 undergoes PS in cells and in vitro (A) Confocal microscopy images of HEK293T cells expressing GATA3WT-mEGFP (green). Nuclei were stained with DAPI (purple). Scale bar, 5 μm. (B) Line-scan intensity profiles of GATA3WT-mEGFP and DAPI signals along the yellow arrow in (A). (C) Time-lapse snapshots of fluorescence recovery after photobleaching (FRAP) in HEK293T cells expressing GATA3WT-mEGFP. Scale bar, 2 μm. (D) FRAP recovery kinetics of GATA3WT-mEGFP condensates (n = 3). t1/2, half-recovery time. (E) Representative snapshots of HEK293T expressing GATA3WT-mEGFP with 5% 1,6-hexanediol treatment across the indicated time. Scale bar, 2 μm. (F) Representative images of 10 μM purified MBP-GATA3WT protein solution in tubes and under microscope with or without TEV protease treatment. Scale bar, 20 μm.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies GATA3 abcam Cat# ab199428 GAPDH abcam Cat# ab8245 H3K27ac abcam Cat# ab4729 H3K9me3 abcam Cat# ab8898 H3 abcam Cat# ab1791 suv39H1 abcam Cat# ab283262 Anti-RNA polymerase II (phospho S2) abcam Cat# ab193468 Anti-KAT3B / p300 abcam Cat# ab275378 Rabbit polyclonal to GFP abcam Cat# ab290 Rig-I/DDX58 Rabbit mAb ABclonal Cat# A22478 IRF-3 Rabbit mAb Cell Signaling Technology Cat# 4302 Phospho-IRF-3 (Ser386) Rabbit mAb Cell Signaling Technology Cat# 37829 Phospho-IRF-3 (Ser396) Rabbit mAb Cell Signaling Technology Cat# 4947 HA-Tag Rabbit mAb ABclonal Cat# AE105 TBK1/NAK Rabbit mAb ABclonal Cat# A3458 Donkey anti-goat IgG-HRP abcam Cat# ab6885 Phospho-TBK1/NAK (Ser172) Rabbit mAb Cell Signaling Technology Cat# 5483 Anti-KMT1E/SETDB1 abcam Cat# ab300573 Estrogen Receptor alpha Santa Cruz Cat# sc-73562 PML Santa Cruz Cat# sc-71910 Anti-SC35 abcam Cat# ab204916 Mouse monoclonal [IH10] to Coilin abcam Cat# ab87913 Bacterial and virus strains E. coli DH5α Lab stored N/A E. coli BL21/DE3 Lab stored N/A Lentivirus Lab stored N/A Chemicals, peptides, and recombinant proteins IPTG Sigma-Aldrich Cat# I6758 PMSF Sigma-Aldrich Cat# 329-98-6 LipofectamineTM 2000 Thermo Fisher Scientific Cat# 11668027 Puromycin Sigma-Aldrich Cat# 540222 Chaetocin MCE Cat# 28097-03-2 5-Azacytidine MCE Cat# 320-67-2 β-Estradiol MCE Cat# 50-28-2 Cycloheximide MCE Cat# 66-81-9 1,6-Hexanediol aladdin Cat# 629-11-8 X-tremeGENETM HP DNA Sigma-Aldrich Cat# 6366244001 Recombinant MBP-GATA3-WT protein This study N/A Recombinant MBP-GATA3-N protein This study N/A Recombinant MBP-GATA3-C protein This study N/A Recombinant MBP-GATA3-DBD protein This study N/A Recombinant MBP-GATA3-ΔN protein This study N/A Recombinant MBP-GATA3-ΔC protein This study N/A (Continued on next page) 18 Cell Reports 44, 115702, May 27, 2025
Techniques: In Vitro, Confocal Microscopy, Expressing, Staining, Fluorescence, Purification, Microscopy