Stellaris smFISH probes targeting , an ortholog of the human solute carrier transporter SLC50A1, are shown in red (Cal Fluor 610), DAPI in blue, and PGL-1::GFP, a marker for P granules, in green. A) transcripts are expressed in Z1/Z4 cells during embryogenesis (arrowheads). The embryo shows Z1 and Z4 migrating to Z2/Z3 P granule cells (marked with PGL-1::GFP in green). B) A dissected proximal gonad of a young adult capturing the ovulation of a recently fertilized zygote (arrow) as it passes through the expressing spermatheca (red). Proximal oocyte marked with arrowhead. C) Sperm nuclei (blue) within the (red) spermatheca. scale = 20µ The experiment confirms and extends enrichment in cell-specific Z1/Z4 mRNA profiling experiment reported by Kroetz & Zarkower 2015.
Reagents
The Stellaris smFISH probe contains 44 oligos labeled with Cal Fluor 610 (sequences available upon request). Staining was performed as described by Ji and van Oudenaarden (2012). In short, PGL-1::GFP embryos and worms were fixed with formaldehyde and ethanol. Hybridization was done in the dark at 37°C for four hours. Cal Fluor 610 coupled probes were designed with the Stellaris Probe Designer from Biosearch Technologies.