Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 (Revised) - Supplemental Figure 1. ARPC1 Copy Number Analysis A Southern analysis of P. patens genomic DNA digested with both single and non-cutters (Nde I, Bgl II, EcoR V) within the known ARPC1 genomic sequence. The Schematic illustrates the 3.1 Kb fragment of ARPC1 utilized to probe P. patens genomic DNA. The sites within the probe for the single cutter enzymes (Msc I and Xcm I) are shown. Low stringency wash conditions optimized to detect at least 70% sequence identity were used. The presence of more than one band in the non-cutter lanes and more than two bands in the single cutter lanes demonstrates the presence of an additional ARPC1 gene in the genome.
• Supplemental Figure 2 - Supplemental Figure 2. The NLS-GFP-GUS Locus is Disrupted in arpc1 lines A Southern analysis of P. patens genomic DNA from indicated lines digested with the Nhe I, a non-cutter within the NLS-GFP-GUS locus. A 0.6 Kb fragment of NPTII, the selection cassette for the NLS-GFP-GUS locus, was used as a probe. A wild type negative control and NLS4 and gfp RNAi positive controls are shown. The lack of bands in the arpc1 lanes shows the complete loss of the NPTII sequence. Due to the high frequency of homologous recombination in P. patens, we propose integration of the ARPC1 RNAi construct at this locus.