XB-IMG-155940
Xenbase Image ID: 155940
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Fig. 6. : Rcsd1 functionally interacts with CapZ in the cytosol. A. Schematic overview of used murine Rcsd1 and deletion constructs. Rcsd1 contains a CPI (capping protein interaction) and a NLS (nuclear localization signal) motif. Numbers indicate amino acids in the Rcsd1 protein. B. Amino acid sequence of murine CPI and NLS motifs. Numbers indicate amino acids in the Rcsd1 protein. C. Transfection of the full-length EGFP-Rcsd1 fusion construct revealed that Rcsd1 is localized in the nucleus and cytosol (upper row). Deletion of CPI (middle row) or NLS (lower row) in Rcsd1 leads to a cytosolic localization. Scale bar: 100 μm. D-E. Split YFP complementation assays. Murine full-length Rcsd1 and Rcsd1δNLS (labelled as δNLS) interact with CapZa and CapZb whereas Rcsd1δCPI (labelled as δCPI) does not. Scale bars D, E: 10 μm. F. Rcsd1 MO was unilaterally injected along with different murine Rcsd1 constructs and the expression of the cardiac markers tnni3, actc1, myh6 and alcam was monitored at stage 28 (ventral views). Reduced marker gene expression upon loss of Rcsd1 (red arrowheads) was rescued by co-injection with the full-length murine Rcsd1 construct (Rcsd1, dark grey arrowheads) and Rcsd1δNLS (dark blue arrowheads). Rcsd1δCPI however (orange arrowheads) was not able to restore marker gene expression. G. Quantitative presentation of data shown in F. n, number of independent experiments; N, number of analysed embryos. Error bars indicate standard error of the means (s.e.m.). n.s., not significant, *, p≤0.05; **, p≤0.01; ***, p≤0.001; calculated by a non-parametric Mann-Whitney rank sum test. Image published in: Hempel A et al. (2017) Copyright © 2017. Image reproduced with permission of the Publisher, Elsevier B. V.
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