As a framework for future transcriptomic investigation of the evolution of these highly derived parasites, this study used the filtered set of transcripts to mine for taxonomically restricted genes as well as genes with non-cnidarian homologs, describe for the first time putative venom homologs, and characterize proteins related to extracellular communication.
mTurquoise and SYFP2 were imaged using the filter set used for Cfp and Yfp, respectively.
The PLA signal was quantified by a FACScan on LSRFortessa (Becton Dickinson, Franklin Lakes, NJ) using the filter set for PE.
Images were not subjected to deconvolution and were captured at 520 550 nm using the filter set # 89006 (Chroma Technology).
mCherry/RFP expression was imaged using excitation at 556/20 nm and emission between 590 and 650 nm using the filter set 51019 (Chroma, Bellow Falls, VT, USA).
FM4-64 samples samples were imaged using a Spot RT CCD camera mounted on a Nikon Eclipse E800 fluorescence microscope using the filter set 532-587 nm excitation, 595 nm long pass dichroic mirror, 608-683 nm emission.
For auramine O staining, pollen grains were suspended in 0.1% auramine O and 50 mM Tris-HCl, pH 7.5 and observed under a Zeiss LSM-510 laser-scanning confocal microscope using the filter set suitable for FITC.
Ncc is the inverse amplitude of the cross-correlation curve, and the cross-talk between channels, Q, was estimated to be approximately 5% for the GFP and mCherry probes using the filter sets described above.
Chlorophyll autofluorescence, MitoTracker and GFP fluorescence of the cells were dissected using the filter sets U-MWSG2 (Olympus), 41007 and 41020 (Chroma Technology Corp, Rockingham, VT, USA), respectively.
Using the filtered data sets we follow a more thorough analysis of the coexpression distributions with respect to the parameters r and N.
