XB-IMG-152678
Xenbase Image ID: 152678
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Figure 1. Actinin tension sensor in cultured cell and embryo.(a) Schematic diagram of tension sensor. The FRET domain was inserted between SR1 and SR2. ABD: actin binding domain. SR1-4: spectrin repeat domain. CLD: calmodulin-like domain. (b) High FRET control. The FRET domain was attached to the C-terminal of actinin with two linking amino acid residues. (c) Mutant non-fluorescent constructs. To break EGFP fluorescence, the 66th tyrosine of the EGFP domain was replaced with leucine. To break mCherry fluorescence, the 72nd tyrosine of the mCherry domain was replaced with leucine. Tyr66 of EGFP and Tyr72 of mCherry compose chromophores. (d–i) Acceptor images (left), donor images (center), and corrected FRET index images (right) of HEK cells (d–f) and Xenopus ectoderm (g–i) expressing ActTS-GR (d,g), hiActTS-GR (e,h), and a pair of ActTS-GR non-fluorescent mutants (f,i). (j,k) Quantification of the FRET index in HEK cells (j) and ectoderm (k). We measured >7 cells and >6 embryos per construct and values are average ± SD. (l) Schematic of the tension sensor in a cell. ActTS-GR makes an antiparallel dimer and bridge between actin filaments. Tension on actin filaments stretches FRET domain of ActTS-GR. Scale bars = 10 μm in f and 50 μm in i. Image published in: Yamashita S et al. (2016) Copyright © 2016, Macmillan Publishers Limited. 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 |