Extended Data Fig. 4: Cytoplasmic localization of TFL1 in Arabidopsis.

a,b, Localization of TFL1-GFP in developing tfl1-20 gTFL1-GFP seeds stained with DAPI at 4 (a) and 5 (b) days after pollination (DAP) by cryosectioning. TFL1-GFP signal in the syncytial (a) and cellularized (b) peripheral endosperm does not co-localize with DAPI staining. The inset in each panel in (a) or (b) shows one enlarged nucleus of the syncytial or cellularized peripheral endosperm, respectively. DAPI, fluorescence of 4,6-diamino-2-phenylindole; BF, bright-field image; Merge, merge of GFP, DAPI and BF images. Scale bars, 50 µm. c, Analysis of TFL1-3HA cytoplasmic localization by immunogold electron microscopy using anti-HA antibody in the syncytial peripheral endosperm of tfl1-20 gTFL1-3HA seeds at 4 DAP. The lower panel shows a higher magnification of the area within the box indicated in the upper panel. Arrows indicate gold particles. There are no detectable gold particles in the nucleus. CV, central vacuole; iCW, cell wall of ii1; ii1, the innermost cell layer of the inner integument (endothelium cell); N, nucleus; Nu, nucleolus; PSV, protein storage vacuoles; SPE, syncytial peripheral endosperm. Scale bars, 2 µm. d, Subcellular localization of TFL1-GFP in the cells of a tfl1-20 gTFL1-GFP inflorescence meristem. Merge, merge of GFP and bright field images. Scale bar, 10 μm. The experiments in a-d were repeated three times independently with similar results.