a, Wild-type (Col-0) and BIN2:BIN2-GFP;Col-0 plants (lines 21 and 26), grown for 4 weeks in soil with corresponding BIN2 gene expression in seedlings measured by qRT–PCR. n = 3 biologically independent experiments. Scale bars, 2.5 cm b, Schematic representation of the tDNA insertions in the atsk12 mutant (GABI-410D09). The insertion site is annotated with the flanking sequence and corresponding insertion site or inserted nucleotide in green. Primers used are depicted in the scheme and listed in Supplementary Table 2. Wild-type (Col-0) and ATSK12:ATSK12-GFP;atsk12 complemented plants (lines 2, 3 and 14) grown for 2 weeks in soil with the corresponding ATSK12 gene expression in seedlings measured by qRT–PCR. n = 3 biologically independent experiments. All qRT–PCR were carried out with seedlings at 5 d.p.g. Transcript levels were normalized to the ACTIN2 gene expression. c, In vitro phosphorylation of YDA and SPCH by ATSK12. All experiments were repeated independently three times. Box plots show the first and third quartiles, split by the median (line) and mean (cross). For blot source data, see Supplementary Fig. 1.