Extended Data Fig. 4: Characterization of different DNA Ligases, Pol1A and Pol1B.
From: PEN1 catalyses RNA primer removal during plastid DNA replication in maize
a,b,c, Subcellular localization of LIG4-GFP, LIG6-GFP, and LIG1-GFP in N.benthamiana leaf epidermal cells. Chl, chlorophyll autofluorescence (magenta). DAPI was used to stain the nucleus. Scale bars: 10 μm. More than six replicates, consistent results. d, High-resolution image of LIG1-TDT in the chloroplast. DAPI was used to stain the nucleus and chloroplast nucleoids. The PEN1-GFP was used for co-localization analysis. Scale bars: 2 μm. More than six replicates, consistent results. e, Purity analysis of LIG1 (253-909aa), Pol1A (257-1052aa), and Pol1B (245-1039aa). SDS-PAGE analysis of the LIG1, Pol1A, and Pol1B. M represents the protein molecular Marker. Three replicates, consistent results. f, Subcellular localization of Pol1A-GFP in N.benthamiana leaf epidermal cells. Scale bars: 10 μm. More than six replicates, consistent results. g, High-resolution image of Pol1A-TDT in the chloroplast. The PEN1-GFP was used for co-localization analysis. Scale bars: 2 μm. More than six replicates, consistent results. h, Subcellular localization of Pol1B-GFP in N.benthamiana leaf epidermal cells. Scale bars: 10 μm. More than six replicates, consistent results. i, High-resolution image of Pol1B-TDT in the chloroplast. The PEN1-GFP was used for co-localization analysis. Scale bars: 2 μm. More than six replicates, consistent results. j, Phylogeny of plastid Pol1A and Pol1B homologs in representative plant taxa. The simplified phylogenetic tree illustrates the relationships among plant species from which Pol1A and Pol1B homologs were identified. Given the high protein similarity between Pol1A and Pol1B, only Pol1A was selected for phylogenetic analysis.
