Xenbase Image ID: 145573

Fig. 6. DHFR inhibition results in increased apoptosis and DNA damage. (A) Schematic of the experimental design. (B) Immunohistochemistry of cleaved caspase-3 (green) in transverse sections of the face from embryos treated with DMSO (i), or MTX (ii,iii) or injected with control morpholino (CMO)(iv) or DHFR morpholinos (DHFR MO) (v,vi). Images are all at the same magnification (scale bar=100 μm) and counterstained with propidium iodide. (C) Imagestream flow cytometry to analyze cell morphology. (Ci) Schematic showing cellular measurements used in the analysis. Cii) Bar graph showing average perimeter (Per.) and Aspect ratio (Aspect) (major/minor axis). Statistical significance designated by asterisks (p values≤0.05). (Ciii) Representative images of single cells acquired by the ImageStream. (D) Comet assay for DNA damage. (Di) Representative cell to show the comet tail region where fluorescence was quantified. (Dii) Bar graph showing quantification of fluorescence in comet tails from cells isolated in two experiments. Statistical significance is designated by an asterisk (n=40 cells, p value=6.8E–06). (Diii,iv) Representative images of cells from comet assay. Scale bar=50 μm. Image published in: Wahl SE et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, Elsevier B. V. Gene Synonyms Species Stage(s) Tissue casp3.2.L Apopain, CASP-3, caspase 3, caspase-3, xCPP32 X. laevis Sometime during NF stage 29 and 30 to NF stage 31 brain eye Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + Methotrexate Hydrate + NF35/36 (immunohistochemistry) fig.6.b. ii, iii Anatomical Phenotype increased apoptosis in head Xla Wt + dhfr MO + NF35/36 (immunohistochemistry) fig.6.b. v, vi Anatomical Phenotype increased apoptosis in head Larger Image Printer Friendly View

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