Supplementary Figure 8: The effect of knockdown of class I BPC or C1-2iD Zn-finger TF family in the hypomorph clf^R mutant.

(a) Leaf curling (inset) and flowering time in wild type (WT), the weak clf^R mutant, class I BPC knockdown in clf^R, C1-2iD ZnF knockdown in clf^R and in clf-50 RNA null mutant. Photographs show plants 6 weeks after germination. Scale bar, 1cm. (b) Leaf curling phenotype of independent BPC^KD clf^R (top) and ZnF^KD clf^R (bottom) transgenic lines. Photographs show plants 4 weeks after germination. Scale bar, 1cm. (c) Quantification of the phenotypes in (b). Box and whisker plot with median (red line, n = 15 plants), upper and lower quartile (box edges), and minima and maxima (whiskers). Letters above boxes indicate significantly different groups (P < 0.05 based on Kruskal–Wallis test with Dunn's post hoc test). (d) De-repression of Polycomb target gene expression (AG, SEP3, LEC2) in the genotypes in (b). Expression is shown relative to the parental line (clf^R). The housekeeping gene EIF4 served as control. (e) The abundance of the CLF mRNA relative to the wild type as measured by qRT-PCR. CLF levels are unchanged in the BPC^KDclf^R and the ZnF^KDclf^R plants shown in (b) compared to the parental line (WT). Shown are mean ± s.e.m. of one experiment for three independent transgenic lines (d,e).