XB-IMG-130465
Xenbase Image ID: 130465
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Figure 1. Time dependent induction of the early nephrogenic transcription factors in retinoic acid and activin A treated animal caps. (A) Animal caps were cut at the late blastula stage 9 [36] and incubated for 1.5 or 3 hours in retinoic acid (10-4M) and activin A (10 ng/ml). For 5, 7 and 13 hours treatments retinoic acid and activin A was replaced after three hours with Steinberg's solution [15]. The mRNA levels were analysed by quantitative RT-PCR using the primers given in Additional file 2. The standard deviations from four independent animal cap pools (N = 30) are given and alterations are indicated by an upward arrow, if all four induction values were higher than one for a given probe. To detect osr2 transcripts primers targeting osr2B, the splice variant predominantly expressed in Xenopus laevis (compare Figure 2Bs) were used for two animal cap pools analyzed after 3, 5 and 7 hours and three after 13 hours In all other experiments primers targeting both splice variants were used. (B) Animal caps cut at the late blastula stage 9 were incubated with or without cycloheximide (CHX, 5 μg/ml) for 30 min and then stimulated for 1.5 hours in retinoic acid (10-4M) and activin A (10 ng/ml). RNA was quantified from six pools (N = 30) and upward arrows indicate induction for all six experiments. (C) Animal caps were cultured for three hours in retinoic acid (RA, 10-4M), activin A (ActA, 10 ng/ml) or both inducers together (RA+ActA) and then analysed by quantitative RT-PCR. The standard deviations from five (RA) or four (ActA or RA+ActA) independent animal cap pools (N = 30) are given and reproducible induction or reduction is indicated by an upward or downward arrow, if increased or decreased in all samples, respectively. Image published in: Drews C et al. (2011) Copyright ©2011 Drews et al; licensee BioMed Central Ltd. This image is reproduced with permission of the journal 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 |