Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Supplementary figure 1 Analysis of expression levels (A) Semi-quantitative RT-PCR showing the relative expression strength of the transgenic constructs Pro_GL2:ICK1/KRP1, Pro_GL2:ICK1/KRP1¹⁰⁹, Pro_GL2:YFP:ICK1/KRP1, Pro_GL2:ICK1/KRP1¹⁰⁹:YFP, Pro_GL2:GUS:YFP:ICK1/KRP1¹⁰⁹, Pro_UAs:ICK1/KRP1¹⁰⁹ in #254. The expression strength was compared with the endogenous expression of translation elongation factor 1 (EF1). The numbers at top indicate the RT-PCR cycle number. Pro_GL2:YFP:ICK1/KRP1 and pUAS:ICK1/KRP1¹⁰⁹appeared to be slightly stronger expressed than the other transgenes. (B) Western Blot analysis of Pro_GL2:YFP, Pro_GL2:YFP:ICK1/KRP1, and Pro_GL2:ICK1/KRP1¹⁰⁹:YFP misexpressing plants with an antibody against GFP/YFP. As a loading control Ponceau staining of the membrane after protein transfer is shown in the lower panel. From extracts of Pro_GL2:YFP plants a band of approximately 27 KDa was detected matching the calculated size of YFP. No bands could be detected for YFP:ICK1/KRP1. For ICK1/KRP1¹⁰⁹:YFP a band was detected at the expected fusion protein size of approximately 37 KDa, in addition, a faint band appeared at about 27 KDa resembling most likely a degradation product. ICK1/KRP1 is abbreviated as KRP1 marked by an asterisk.
• Supplemental Figure 2 - Supplementary figure 2 Analysis of ICK1/KRP1 movement Confocal-laser-scanning micrograph of a (A) young and (B)old rosette leaf of Pro_GL2:YFP expressing plants. Note that the YFP signal can be observed in the nucleus and the cytoplasm of trichomes and their surrounding cells of young and old leaves. (C) Analysis of ICK1/KRP1 movement in Pro_GL2:YFP:ICK1/KRP1 and Pro_GL2:ICK1/KRP1¹⁰⁹:YFP misexpressing plants in comparison to plants expressing Pro_GL2:YFP as a control. For analysis the ratio of the average YFP intensity of trichome-neighboring cell nucleus to the average YFP intensity of the young trichome nucleus was determined. Whereas the smaller ICK1/KRP1¹⁰⁹:YFP fusion protein appears to move similarly to YFP, the larger fusion of the full length ICK1/KRP1 to YFP is found at lower levels in the nuclei of trichome-neighboring cells in comparison to trichome nuclei. Scale bar in (A) and (B) 50μm. ICK1/KRP1 is abbreviated as KRP1 marked by an asterisk.