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Fig. 3.
Detection of Xenopus P2X4 mRNA in oocytes and surrounding tissues by RNase protection assay. RNase protection assays performed on total cellular RNA (20 μg A and 4 μg B) extracted from defolliculated, devitellinated oocytes (lane 1), oocytes with intact follicle cell layer (lane 2), the transparent vascularized epithelial sac which surrounds each ovarian lobe minus the folliculated oocytes (lane 3) and the entire ovarian lobe with folliculated oocytes at various stages I–VI and the vascularized sac surrounding them (lane 4). 200 000 cpm of each riboprobe ((a) the 318 nucleotide Xenopus P2X4 riboprobe (asterisk in lane P; protects 256 nucleotides), (b) the 220 nucleotide Xenopus histone H4 riboprobe (asterisk in lane P; protects multiple sizes of RNA) was used in the reactions and the gels were exposed to X-ray film for 25 h (a) or 3 h (b). The riboprobes were used at 2000 cpm/lane as size markers (lane P, size in nucleotides as indicated). |
