Journal: Cellular and Molecular Life Sciences
Article Title: APC mutations disrupt β-catenin destruction complex condensates organized by Axin phase separation
doi: 10.1007/s00018-023-05068-0
Figure Lengend Snippet: Axin is involved in the β-catenin nuclear translocation and transcriptional function as a droplet. A Confocal images of intracellular Axin (red) and β-catenin (green) in CRC cells. Line four: representative images of 3D reconstitution using by Imaris software. Line five: section drawing of 3D reconstructed confocal images by Imaris software. The tool, Clipping Plane, which allows to re-slice in any chosen rotation ( https://imaris.oxinst.com/ ), uncovers hidden regions by re-slicing the image data in a different orientation. B Representative images of Axin (red) and nuclear (DAPI, blue) expression after transfection of NLS-mutated Axin in HEK293T and SW480. C Cells were treated with or without Leptomycin B (LMB, 200 nM, 24 h). Nuclear and cytoplasmic protein extraction experiment was further performed to measure the cellular localization of Axin in SW480 cells after transfection of NLS-mutated Axin. D The mRNA levels of Wnt pathway-specific downstream target gene (Axin2) were examined by RT-PCR in SW480 cells expressing wild-type Axin or NLS-mutated Axin. Error bars, mean ± SD; **, p < 0.01. E Analysis of TOP/FOP reporter activity in SW480 cells transfected with wild-type Axin or NLS-mutated Axin. Error bars, mean ± SD; **, p < 0.01. F Representative images of β-catenin (green) and nuclear (DAPI, blue) expression after transfection of β-catenin in Axin KO SW480 or wild-type SW480 cells. G Representative fluorescence images of live SW480 and HEK293T after transiently expressing EGFP-tagged Med1-IDR and pScarlet-tagged Axin
Article Snippet: Line five: section drawing of 3D reconstructed confocal images by Imaris software.
Techniques: Translocation Assay, Software, Expressing, Transfection, Protein Extraction, Reverse Transcription Polymerase Chain Reaction, Activity Assay, Fluorescence
