Extended Data Fig. 9: CRISPR efficiency and different phenotypes of the p34 mutants.
a, Gene editing efficiency analysis of three random 9-day-old plants of p34iCRISPR-1 and p34iCRISPR-2 after induction with 10 μM β-oestradiol (Est). The sequencing results were analysed with the online software Inference of CRISPR Edits (ICE) (https://ice.synthego.com/#/). b, P34-GFP localization in roots of 5-day-old p35S:gP34-GFP/p34iCRISPR-1 plants grown on DMSO and 10 μM Est. Arrows indicate the remaining P34-GFP signal. Scale bar, 10 µm. c, Protein levels of AP1G, AP2A and AP2S in p34-3 mutants analysed by immunoblotting with α-AP1G, α-AP2S, α-AP2A, α-CHC, α-TPLATE and α-Tubulin, The experiments were repeated three times. One representative experiment is shown. d, Quantification of protein levels shown in (c). The protein level was normalized to tubulin. e, Confocal images of the 5-day-old pP34:gP34-GFP/p34-1(-/-) (line #12) root cells stained with FM4-64 (2 μM, 40 min). Scale bar, 10 µm. The imaging was repeated three times. One representative experiment is shown. f, FM4-64 uptake in the p34-3 mutants. g, Fluorescence intensity ratio of the relative intracellular-to-plasma membrane (PM) FM4-64 signal. All the individual value were plotted and the center line represents the median. ***P ≤ 0.001 (one-way ANOVA test); ns, not significant. The P values versus the Col-0 control for p34(Δ/-) = 0.0004, p34(Δ/Δ) = 0.6980 and ap2m-2 < 0.0001. n = 30, n number of cells analysed, Scale bar, 10 µm.
