XB-IMG-175624
Xenbase Image ID: 175624
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FIGURE 3. YWHAZ(S230W) rescues embryonic defects induced by DN-FGFR1 more efficiently than YWHAZ. (A) Expression of DN-FGFR1 (25 pg or 50 pg) in the dorsal marginal zone of early embryos results in gastrulation defects, characterized by reduced body length and exposed mesendoderm (“open back”). Co-expression of either YWHAZ or YWHAZ(S230W) (250 pg) with DN-FGFR1 leads to phenotypic rescue, but the S230W variant is more efficient in restoring body length and embryo morphology. The scatter plot of the body length of the embryos showed that the S230W variant rescued the defects more efficiently than YWHAZ. Student’s t-tests were performed in a pairwise fashion and the results revealed significant differences in the samples we compared. This experiment has been repeated four times, with 12 to 18 treated embryos in each experiment. Since the embryos were collected at slightly different stages and the body length was thus different, scatter plot was made for the embryos for each experiment separately. In all the experiments, there was a significant difference between YWHAZ and YWHAZ(S230W) in phenotype rescue. (B) DN-FGFR1 interferes with the expression of the pan-mesodermal marker brachyury, and YWHAZ(S230W) rescues marker expression to a greater extent compared to wild type YWHAZ. The experiment was repeated 3 times. For control embryos, 1/44 embryos showed a gap in the brachyury ring, and 33/38 embryos injected with DN-FGFR1 RNA had a big gap in the brachyury domain. Expression of YWHAZ and YWHAZ(S230W) led to partial rescue of brachyury expression, resulting in similar brachyury gap in 24/38 and 13/38 embryos, respectively. These numbers are indicated in the panel. (C) Ectopic expression of YWHAZ weakly expands, whereas ectopic expression of YWHAZ(S230W) strongly expands, the domain of brachyury in gastrulating Xenopus embryos. The experiment has been repeated 3 times. The expansion of brachyury domain was observed in 16/30 embryos expressing YWHAZ and 24/30 embryos expressing YWHAZ(S230W). The doses of RNAs used in this panel are 1 to 2 ng. The red color in panels B and C are staining of beta-galactosidase-expressing cells with the chemical red-Gal to mark the injected cells. Image published in: Popov IK et al. (2019) Copyright © 2019 Popov, Hiatt, Whalen, Keren, Ruivenkamp, van Haeringen, Chen, Cooper, Korf and Chang. 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
Image source: Published
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