Figure 2: Cytoplasmic AFB1 regulates rapid root growth.
A) Subcellular localization of AFB1-NLS-mCitrine and AFB1-NES-mCitrine (yellow) in afb1–3; roots stained with propidium iodide (magenta). Scale bars = 100 μm.
B) Root growth response (growth of treated roots normalized to growth of the respective mock-treated roots) of Col-0, afb1–3, gAFB1 (afb1–3) and gAFB1-NLS/NES #1, #2 (afb1–3) to IAA (10 nM; 20 minutes).
C) The ccvTIR1-E12K-mScarlet (Col-0) protein (green) localizes to nuclei. Stained with propidium iodide (magenta), scale bar = 50 μm.
D) Root growth rate of ccvTIR1-mScarlet (Col-0) and ccvTIR1-E12K-mScarlet (Col-0) in mock or 500 nM cvxIAA treated seedlings.
E) Subcellular localization of AFB1, AFB1(K8E), TIR1, and TIR1(E12K)-GFP in Col-0 Arabidopsis protoplasts (left) and quantification of relative nuclear localization (right), calculated as the ratio of nuclear fluorescence to total cell fluorescence.
Scale bar = 50 μm.
F) Root growth response of Col-0, afb1–3, cul1–6 and cul1–7 to IAA (10 nM; 20 minutes).
G) Expression pattern and subcellular localization of AFB1-mCitrine in Col-0 and cul1–6 background. Scale bar = 100 μm. Boxplots in B,D,E,F represent the median and the first and third quartiles, and the whiskers extend to minimum and maximum value; all data points are shown as dots. Letters indicate statistical differences according to Ordinary one-way ANOVA coupled with Tukey’s multiple comparison tests (p<0.05).
