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. 2018 Nov 13;9(6):e02135-18. doi: 10.1128/mBio.02135-18

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Copyright © 2018 Zhao et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 4 — The PAS domain is important for the nuclear enrichment of Pas3 and its full function in regulating filamentation. (A) Diagram of Pas3 alleles with mutation or deletion in the core of PAS domain of Pas3. ClustalW multiple alignment of the PAS domain amino acid sequences in Per (AAH72458.1), Arnt (NP_001284648.1), Sim (AAB62395.1), HIF1α (NP_001521.1), Ahr (NP_001612.1), and Pas3 was performed. G72 and Y73 residues in Pas3 are indicated with stars. (B) Western blotting of mCherry-tagged Pas3 alleles probed with an anti-mCherry antibody. (C) Self-filamentation assay of WT, pas3Δ, pas3Δ/PAS3, pas3Δ/PAS3^G72A, pas3Δ/PAS3^Y73A, and pas3Δ/PAS3^CORE⁻ strains. (D) Fluorescence of mCherry-tagged Pas3 alleles (images at bottom) and the quantitative analysis of their subcellular localization in the population. DAPI staining was used to visualize the nucleus (black-and-white images in the middle). The relative fluorescence intensities determined for DAPI and mCherry were plotted along the indicated lines. The percentages of cells with nucleus-enriched mCherry signal and the number of cells measured are indicated on the top right corner.