(A) Sqt-/Cyc-/sec-EGFP and Acvr2b-mCherry constructs. S, signal peptide; Pro, pro-domain; Mat, mature-domain; ECM, extracellular and transmembrane domain. Red arrows indicate convertase cleavage sites. Sqt signal peptides and pro-domain were used in sec-EGFP constructs. (B) Injection procedure. (C) Representative image of an injected embryo at 30% epiboly stage showing the expression patterns of the fusion proteins. Scale bar represents 50 μm. (D,E,F) Representative auto-correlation (ACF) and cross-correlation functions (CCF) and fits. (G,H,I) Individual Ln(Kd) frequency histogram and Gaussian fitting (red curve). Inset, concentration plot and linear regression (red line). X axis, concentration of bound protein (Cgr (x10-9 M)); Y axis, products of concentrations of free proteins (Cg x Cr (x10-16 M)). n = number of data points (number of embryos).
DOI:
http://dx.doi.org/10.7554/eLife.13879.006