Fig. 6: The basic domain of bridgin has a property to interact with DNA nonspecifically in vitro and in vivo.
From: Bridgin connects the outer kinetochore to centromeric chromatin
a Proteins eluted from TBZ-treated cells after FLAG-IP of Bgi1FL expressing SHR879, Bgi1BDΔ expressing SHR880, and a FLAG control strain SHR942 expressing Bgi1 were separated on a gradient PAGE gel and silver-stained. Blue arrows mark the bait proteins. b Electrophoretic mobility shift assay (EMSA) samples were separated on a PAGE gel and stained with GelRed for visualization. c Chromatin-bound proteins (green) colocalize with the nuclear marker histone H4-mCherry in metaphase, while free nuclear proteins diffuse into the cytoplasm following the entry into mitosis. d Visualization of GFP-Bgi1 when expressed under the native (SHR847, BGI1::BGI1-V5-GFP) or an overexpression (OE) GAL7 promoter (SHR858, BGI1::GAL7p-GFP-BGI1). Outer kinetochore protein Ndc80-mCherry was used to mark the kinetochore. e Visualization of bridgin OE constructs. The expression of OE constructs of Bgi1FL (SHR895, SHR832::H3p-GFP-BGI1FL), Bgi1FDΔ (SHR920, SHR832::H3p-GFP-BGI1FDΔ), Bgi1BDΔ (SHR921, SHR832::H3p-GFP-BGI1BDΔ), Bgi1FD (SHR922, SHR832::H3p-GFP-BGI1FD), Bgi1USD (SHR923, SHR832::H3p-GFP-Bgi1USD) and Bgi1BD (SHR924, SHR832::3×FLAG-GFP-BGI1BD), and GFP-PCNA control (SHR854, BGI1, CNVY121::H3p-GFP-PCNA) was visualized in G2 and M phases. Histone H4-mCherry was used to mark chromatin. Scale bar, 3 μm. Source data are available as a Source Data file.
