Xenbase Image ID: 137976

Figure 1 FGFR-VT− expression is spatially and temporally restricted during blastula stage. A, schematic illustrating the 261-nt probe that corresponds to the sequence of the VT+ isoform as well as the VT+ and VT− protected fragments produced by RNase digestion for the analysis shown inB. Digestion of probe:VT+ hybrids results in a 162-nt protected fragment, while digestion of probe:VT− hybrids results in digestion of the 6-nt single strand loop encoding the VT (black box), producing two protected fragments of 107 and 49 nt. The size in nt is listed below each fragment.B, RNase protection analysis of FGFR-VT+ and FGFR-VT− mRNA in Xenopus blastulae. Stage 8 blastulae were dissected into animal, vegetal, and marginal zone regions (as illustrated in the schematic diagram shown above lanes 5–7), as described (14). Total RNA was isolated from each region, and RNase protection analysis was performed using a 32P-labeled 261-nt probe, as in Ref. 7. A representative experiment is shown. Lane 1, probe;lane 2, digested probe; lanes 3–7, protected fragments from in vitro transcribed FGFR-VT+ cRNA, in vitro transcribed FGFR-VT− cRNA, marginal zone cells (M), animal cells (A), and vegetal cells (V), respectively. The positions of the undigested probe (arrow), the VT+ protected fragments (square brackets), and VT− 107-nt protected fragment (arrow) are indicated; the 49-nt VT− protected fragment is not shown. C, RT-PCR analysis of the VT+ and VT− temporal expression pattern in marginal zone cells. Blastula stage embryos were collected at the following postfertilization times: 4.5 h (stage 7), 5.0 h (stage 8), 5.5 h (stage 8), and 6.0 h (stage 8). Marginal zones were dissected, and total RNA was isolated as inB. RT-PCR was performed as described under “Experimental Procedures,” and the VT+ and VT− products, which differ in size by 6 nt, were analyzed by autoradiography on a 6% polyacrylamide, 6M urea sequencing gel. A representative experiment is shown. Amplification products of VT+ cDNA (lane 1) and VT− cDNA (lane 2) mark the position of the VT+ and VT− products (arrows) representing the two isoforms in the marginal zone cells (lanes 3–6). Each marginal zone sample was also amplified using primers for histone (H4), as an input control, and for elongation factor 1-α, as a measure of zygotic transcription.Dev Time, development time. Image published in: Paterno GD et al. (2000) Copyright © 2000. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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