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. 2023 May 15;26(6):106880. doi: 10.1016/j.isci.2023.106880

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© 2023 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

UFO mediates LFY degradation and requires the UFO F box domain and lysine residues in the IDR of LFY

(A) Confocal microscopy images of lfy-12/LFY::LFY-GFP (left) and ufo-2/lfy-12/LFY::LFY-GFP (right) flowers. Scale bar = 60 μm.

(B) Quantification of fluorescence intensity from flowers as shown in (A). Each data point represents the average of fluorescence intensities from 5 LFY-GFP-positive regions of interest in independent flower buds. Independent plants were used for each data point (n = 26). Statistical significance was determined using a two-tailed Student’s t test.

(C) Total LFY protein levels in N. benthamiana leaves transiently expressing 35S::LFY-mCit or coexpressing 35S::LFY-mCit and 35S::UFO-mCh. Ponceau red staining (PS) serves as loading control.

(D) Quantification of LFY-mCit protein levels in N. benthamiana leaves as shown in (C). Results are shown as mean +/− standard error (n = 3).

(E) LFY and UFO mRNA accumulation in N. benthamiana leaves expressing 35S::LFY-mCit or coexpressing 35S::LFY-mCit and 35S::UFO-mCh.

(F) Bimolecular functional complementation of LFY and UFOΔF-box in N. benthamiana leaves. The absence of interactions between LFY and the UFO F-box domain serves as negative control. Scale bar = 10 μm. The overlays between mCitrine fluorescence channels and the transmission view are shown at the bottom. Arrows indicate examples of cytoplasmic foci.

(G) Total LFY-mCit protein levels in N. benthamiana leaves transiently expressing 35S::LFY-mCit or coexpressing 35S::LFY-mCit and 35S::ΔF-box-mCh. Ponceau red staining (PS) serves as loading control.

(H) Quantification of LFY-mCit protein levels in N. benthamiana leaves as shown in (G). Results are shown as mean +/− standard error (n = 3).

(I) LFY and UFO mRNA accumulation in N. benthamiana leaves expressing 35S::LFY-mCit or coexpressing 35S::LFY-mCit and 35S::ΔF-box-mCh.

(J) Bimolecular functional complementation of LFY_5KR and UFO in N. benthamiana leaves (left). Interaction between LFY_5KR and LFY is also tested (right). Scale bar = 10 μm. The overlays between mCitrine fluorescence channels and the transmission view are shown at the bottom. Arrows indicate examples of cytoplasmic foci.

(K) Total LFY_5KR-mCit protein levels in N. benthamiana leaves transiently expressing 35S::LFY_5KR-mCit or coexpressing 35S::LFY_5KR-mCit and 35S::UFO-mCh. Ponceau red staining (PS) serves as loading control.

(L) Quantification of LFY_5KR-mCit protein levels in N. benthamiana leaves as shown in (K). Results are shown as mean +/− standard error (n = 3).

(M) LFY_5KR and UFO mRNA accumulation in N. benthamiana leaves expressing 35S::LFY_5KR-mCit or coexpressing 35S::LFY_5KR-mCit and 35S::UFO-mCh.