Journal: Nature Communications
Article Title: Design and evaluation of a tripartite chemogenetic fluorescent reporter for visualizing ternary protein complexes
doi: 10.1038/s41467-025-62241-8
Figure Lengend Snippet: a Constructs for the expression of bJun-pFAST 1-98 , bFos-pFAST 115-125 and H2B-pFAST 99-114 for probing the Fos·Jun·DNA ternary complex formation. b Representative confocal micrographs of HBR-2,5DM-treated HeLa cells co-expressing bJun-pFAST 1-98 , H2B-pFAST 99-114 and either bFos-pFAST 115-125 or bFosΔ(179-193)-pFAST 115-125 . Cells were treated with 10 μM HBR-2,5DM. Identical imaging settings were used for direct comparison. Scale bars 20 μm. Representative confocal micrographs of n > 100 cells from three independent experiments. c Relative nuclear tripartite-split-FAST fluorescence computed by normalizing the nuclear fluorescence intensity of tripartite-split-FAST I F (tripartite-split-FAST) with the fluorescence intensity I F (iRFP670) of co-expressed iRFP670. Each cell is color-coded according to the biological replicate it came from. The solid circles correspond to the mean of each biological replicate. The black line represents the mean ± SD of the three biological replicates. Source data are provided as a Source Data file.
Article Snippet: The plasmid pAG1713 allowing the mammalian expression of H2B-pFAST 99-114 -P2A-bJun-pFAST 1-98 -IRES-HA-mTurquoise2 was constructed by adding H2B sequence to pAG1256 (CMV pFAST 99-114 -P2A-bJun-pFAST 1-98 -IRES-mTurquoise2, which was itself constructed by adding bJun sequence from Addgene vector #22012 pBiFC-bJunVN173 to pAG1184) for expression of a fusion between H2B and pFAST 99–114 .
Techniques: Construct, Expressing, Imaging, Comparison, Fluorescence
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