The Binding of PKL to Targets Is Largely Dependent on PIF3.
(A) Relative expression of cell elongation–related genes in
Col wild-type,
epp1, pif3, and epp1 pif3
mutant plants. The amounts of mRNA were quantified by RT-qPCR, and the relative expression levels were
normalized to that of a UBQ control.
(B)
ChIP-qPCR assay. Data show the relative
enrichment of IAA19, PRE1, and
ACT2 (negative control) promoter fragments upon
precipitation with anti-PKL antibody or the IgG control.
(C) Immunoblot assay showing PKL protein levels in the
pif3 and pifq mutants and
PIF3 overexpression plants. Immunoblotting using an
anti-tubulin antibody served as the loading control.
(D) Relative PKL expression in the
pif3 and pifq mutants. The amounts of mRNA
were quantified by RT-qPCR, and the
relative expression levels were normalized to that of a UBQ
control.
(E)
ChIP-qPCR assay showing the relative
enrichment of IAA19, PRE1, and
ACT2 (negative control) promoter fragments upon
precipitation with H3K27me3 antibody.
For all experiments, the wild-type and various mutant seedlings were grown in
darkness for 5 d. In (A), (B), (D), and
(E), data represent means ± sd of biological
triplicates. Asterisks indicate significant differences from the wild type at P
< 0.05 (*) or P < 0.01 (**) using Student’s
t test.