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FIGURE 4. dhrs3 negatively regulates the synthesis of endogenous atRA. A, log-log plot showing the relationship between different concentrations of atRA standard applied to HEK293T cells and the respective luciferase activity. B, relative concentration of embryonic atRA under different treatment conditions compared with control as measured by cell-based luciferase assay. Overexpression of cyp26a1 and dhrs3 reduced the endogenous atRA level, whereas overexpression of aldh1a2, rdh10, or knockdown of dhrs3 strongly increased atRA concentration in the embryos. The measured concentration of atRA in cell culture is indicated in the plot as mean � S.D. in nM. Readings were taken in triplicate. C, relative concentration of embryonic atRA in aldh1a2-overexpressing embryos and Dhrs3 knockdown embryos, as measured by LC-MS. Relative level of atRA is represented as fold change compared with control reading. Injection dose of each condition per embryo is as follows: 4 ng for cyp26a1; 4 ng for dhrs3 ORF; 30 ng for Dhrs3 MO; 4 ng for aldh1a2; 4 ng for rdh10. Image published in: Kam RK et al. (2013) Copyright © 2013. 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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