Transcript B Supplementary Figure 2

Supplementary Figure 2
4
B
FLK
**
WT
RNAi4
RNAi5
3
2
1
0
untreated
cis-CA
trans-CA
Relative Expression Level
Relative Expression Level
A
2
FLC
**
WT
RNAi4
RNAi5
1
0
untreated
cis-CA
trans-CA
Supplementary FIG. 2. Quantitative RT-PCR analysis of expression levels of
flowering genes in the ZCE1 RNAi lines. Quantitative RT-PCR analysis shows the
transcript levels of FLK (A) and FLC (B) upon cis-CA and trans-CA treatment,
respectively. FLK is a positive regulator of flowering via suppressing the expression
of another flowering gene FLC, while the role of FLC is to repress flowering. (MiHye Lim et al 2004, Scott D. Michaels and Richard M. Amasino 1999). Actin (ACT2)
is used as the internal control, so each tissue’s transcription level was normalized
against actin. WT untreated group was set to 1.0 for comparison. In the untreated
groups, each kind of untreated tissue was compared with that of WT. In the cis-CA
and trans-CA-treated groups, WT and two ZCE1 RNAi lines were compared to each
untreated tissue. ** means 2 or more folds compared with untreated group. The
qPCR condition was set at 95 C for 2 min; 60 cycles of (95 C, 3 sec; 59 C 30 sec);
60-95 C. Primers used for qPCR are listed as the following:
ACT2 (At3g18780)-real time-F: 5’-GAC CAG CTC TTC CAT CGA GAA-3’
ACT2 (At3g18780)-real time-R: 5’-TCT CGT GGA TTC CAG CAG C -3’
FLK (At3g04610) -real time-F: 5’-CAA AAG CCA ACT CGC CAA AT-3’
FLK (At3g04610) -real time-R: 5’-TGA CCA TAG CCT CCT CCA CC-3’
FLC (At5g10140) -real time-F: 5’-AAC GTC GCA ACG GTC TCA TC-3’
FLC (At5g10140) -real time-R: 5’-CGA TCA AGG ATC TTG ACC AGG-3’