Figure 7.
FIT colocalized with PB markers in distinct PBs. Confocal images showing localization of FIT-GFP and PB markers upon coexpression in the nucleus at t = 0 and t = 15 min. (A–C) Coexpression of FIT-GFP with A, PIF3-mCherry, and B and C, PIF4-mCherry, in B, showing a typical pattern with the absence of NBs (∼50% of nuclei), and in C, showing a typical pattern with the presence of NBs (∼50% of cells). When FIT-GFP was coexpressed with PB markers, FIT NBs did not appear at t = 5 min, but instead, FIT-GFP colocalized with PB markers in PBs at t = 15 min. PIF3-mCherry localized predominantly to a single large PB at t = 0 and t = 15 min. FIT-GFP colocalized with PIF3-mCherry in this single large PB at t = 15 min. PIF4-mCherry and FIT-GFP were both homogeneously distributed in the nucleoplasm at t = 0 and t = 15 min, or FIT-GFP colocalized with PIF4-mCherry in PBs at t = 0 and t = 15 min. The same localization patterns were found for PIF3-mCherry and PIF4-mCherry upon single expression (compare with Fig. S3, H–J). Hence, FIT-GFP was recruited to the two distinct types of PIF3 and PIF4 PBs, whereas PIF3 and PIF4 were not recruited to FIT NBs. This suggests that FIT NBs are affected by the presence of PIF3- and PIF4-containing PBs and a connection to light signaling exists. Scale bar: 2 µm. Arrowheads indicate colocalization in PBs. G = GFP; C = mCherry. Fluorescence protein analysis was conducted in transiently transformed N. benthamiana leaf epidermis cells, following the standardized FIT NB analysis procedure. In all examined cells, PIF3 and FIT colocalized fully, while PIF4 and FIT colocalized as indicated in B and C. Representative images from four to six independent experiments are shown.

FIT colocalized with PB markers in distinct PBs. Confocal images showing localization of FIT-GFP and PB markers upon coexpression in the nucleus at t = 0 and t = 15 min. (A–C) Coexpression of FIT-GFP with A, PIF3-mCherry, and B and C, PIF4-mCherry, in B, showing a typical pattern with the absence of NBs (∼50% of nuclei), and in C, showing a typical pattern with the presence of NBs (∼50% of cells). When FIT-GFP was coexpressed with PB markers, FIT NBs did not appear at t = 5 min, but instead, FIT-GFP colocalized with PB markers in PBs at t = 15 min. PIF3-mCherry localized predominantly to a single large PB at t = 0 and t = 15 min. FIT-GFP colocalized with PIF3-mCherry in this single large PB at t = 15 min. PIF4-mCherry and FIT-GFP were both homogeneously distributed in the nucleoplasm at t = 0 and t = 15 min, or FIT-GFP colocalized with PIF4-mCherry in PBs at t = 0 and t = 15 min. The same localization patterns were found for PIF3-mCherry and PIF4-mCherry upon single expression (compare with Fig. S3, H–J). Hence, FIT-GFP was recruited to the two distinct types of PIF3 and PIF4 PBs, whereas PIF3 and PIF4 were not recruited to FIT NBs. This suggests that FIT NBs are affected by the presence of PIF3- and PIF4-containing PBs and a connection to light signaling exists. Scale bar: 2 µm. Arrowheads indicate colocalization in PBs. G = GFP; C = mCherry. Fluorescence protein analysis was conducted in transiently transformed N. benthamiana leaf epidermis cells, following the standardized FIT NB analysis procedure. In all examined cells, PIF3 and FIT colocalized fully, while PIF4 and FIT colocalized as indicated in B and C. Representative images from four to six independent experiments are shown.

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