Supplemental data

1. def-101 def-101 0h 24h 72h 0h 24h 72h 1 0.54 0.57 1 0.63 0.65 1 0.28 0.21 1 1.31 0.67 1 0.58 0.29 1 0.61 0.48 activated target gene EST # repressed target gene EST # 1 1.67 2.83 ns LTP 018_1_01_c16 flavonone 3-beta- hydroxygenase 018_6_07_m18 * * 1 1.85 1.91 1 0.91 0.59 aquaporin 018_5_05_f17 unknown prot. 018_5_08_f14 * 1 1.10 1.33 CER1-like 018_6_08_n20 hyp. protein 018_4_05_m21 * 1 0.69 1.40 PAP 018_6_08_j23 unknown prot. 018_4_06_a13 * * LTP 018_1_01_n17 Ran3 (control) 018_6_06_k11 ns LTP 018_4_11_h12 * 1 0.41 0.49 -tubulin II 018_6_01_o03 NOI 018_5_09_a01 Supplemental data Figure 1. Confirmation of array results for selected DEF target genes from petal stage 3. In total, 21 target genes identified by the def-101 profiling experiments were independently tested by RT-PCR experiments. Array expression data were confirmed for 19 of them. 9 are shown in Figure 6 and here we show the analysis of 12 further target genes. EST clone identifiers are specified. First strand synthesis was conducted from total RNA extracted from def-101 petals at stage 3 cultivated in the permissive temperature (15°C) and shifted for 0, 24 and 72 h to the non-permissive temperature (26°C). PCRs were conducted with 25 cycles or with 28 cycles (*). As a control, Ran 3 was used. For quantification of the data, band strength was normalized relative to the RAN 3 signal as done for Figure 6. -tubulin expression did not decrease steadily. NOI expression decreased upon a reduction of the DEF function instead of an expected increased expression level.