Extended Data Fig. 7: Analysis of PDK1 transgenic lines.
From: The lipid code-dependent phosphoswitch PDK1–D6PK activates PIN-mediated auxin efflux in Arabidopsis
a, Expression of PDK1.1 or PDK1.2 (p35S::Venus-PDK1.1, p35S::Venus-PDK1.2, p35S::mCherry-PDK1.1 and p35S::mCherry-PDK1.2) rescued the growth defects of pdk1.1 pdk1.2. Adult plants (25-day-old) were observed and representative photos were shown. Scale bar, 2 cm. b, Western blot analysis verified the PDK1.1 or PDK1.2 expression (p35S::Venus-PDK1.1 and p35S::Venus-PDK1.2) in pdk1.1 pdk1.2, respectively. Seven-day-old T3 homozygous seedlings were used for protein extraction and subjected to analysis. Upper panel, anti-GFP (1:2000); lower panel, Ponceau staining. c, Western blot verified the PDK1.1 or PDK1.2 expression (p35S::mCherry-PDK1.1 and p35S::mCherry-PDK1.2) in pdk1.1 pdk1.2, respectively. Seven-day-old T3 homozygous seedlings were used for protein extraction and subjected to analysis. Upper panel, anti-RFP (1:2000); lower panel, Ponceau staining. d–e, mCherry-fused PDK1.1 (d) and PDK1.2 (e) localised to both cytoplasm and the basal side of PM. Four-day-old seedlings of p35S::mCherry-PDK1.1 and p35S::mCherry-PDK1.2 were imaged by CLSM. Open arrowheads indicated the basal polar localisation. Scale bar, 10 µm. f–i, Subcellular localisation of Venus-fused PDK1.1N (f, cytoplasm), Venus-fused PDK1.1C (g, both cytoplasm and nucleus), Venus-fused PDK1.2N (h, cytoplasm), and Venus-fused PDK1.2C (i, both cytoplasm and nucleus). Four-day-old seedlings expressing corresponding fusion proteins were imaged by CLSM. Scale bar, 10 µm. n = 4, 2, 3, and 3 biologically independent experiments for (a), (b), (c), and (d) respectively, with similar results obtained.
