Supplementary Figure 7: RNAi-mediated knockdown of class I BPC and C1-2iD ZnF TF families.

(a,b) Class I BPC TF family knockdown by RNAi (BPC^KD, a) or C1-2iD C2H2 Zn-finger TF knockdown (ZnF^KD, b) was assayed in independent transgenic lines in the hypomorph clf^R mutant background. Expression of all genes tested is relative to that in clf^R. Mean ± s.e.m. shown from one experiment and 3 independent lines. In each case, strong knockdown of the targeted TF family is observed (class I BPC and AZF1, AZF3, ZAT6), with more minor effects on distantly related BPC or ZnF TFs (BPC6, At2g26940, At3g46080). The translation initiation factor EIF4 served as qRT-PCR control. (c) Simultaneous knockdown of Class I and C1-2iD C2H2 Zn TFs by RNAi (BPC + ZnF^KD) in the wild type. Shown are 2 independent double knockdown lines analyzed as described in (a, b). (d,e) Phylogenetic tree of the BBR-BPC family of TFs (see also²²) (d) and of select C2H2 Zinc finger TFs (see also²⁰) plus a subset of additional C1-2i C2H2 ZnF proteins in Arabidopsis (e). Branch length is indicated. Light green shading highlights the class I BPC TFs and light purple shading the C1-2iD Zn-finger proteins targeted for knockdown. Triangles point to the genes tested by qRT-PCR in (a-c) or used as controls in BiFC (Fig. 2d,e; BPC6 and ZAT5). Triangle color indicates RNAi targets (green) or distantly related genes (red). Note that the primer pair employed for class I BPC qRT-PCR amplifies all 3 genes in that clade.