Lsm 5 pascal inverted

For the visualization of RBV-eYFP and HYL1-YFP in transgenic plants, roots of 10-day-old seedlings were observed under a Zeiss LSM 5 Pascal inverted confocal microscope (excitation, 488/512 nm/nm; emission, 520/50 nm/nm). For SE-mRuby3 and SE-eCFP, roots of 10-day-old pSE:SE-mRuby3 seedlings were observed under a LeicaSP8 confocal microscope (excitation, 514 nm; emission, 540–600 nm).

VCaP cells (3x10⁶ cells) were plated in 10 cm cell culture dishes and allowed to adhere for 24 h before addition of DMSO vehicle or polyamide stock in DMSO. After 72 h incubation, cells were washed with warm PBS (37°C), gently scraped, and counted. Samples were centrifuged, resuspended at 1x10⁵ cells/mL, and treated according to manufacturer protocol (Trevigen) for neutral electrophoresis. Slides were stained with SybrGreen (Trevigen) and imaged at the Caltech Beckman Imaging Center using a Zeiss LSM 5 Pascal inverted laser scanning microscope equipped with a 5x air objective lens. Overlayed fluorescence and bright field images were obtained using standard filter sets for fluorescein. Images were analyzed using Comet IV software (Perceptive Instruments Ltd) with 200–600 comets measured per sample. A random sampling of 200 comets per condition was used for two-way analysis of variance (ANOVA) analysis (Prism software, GraphPad) of three biological replicates.