Figure 4. The distribution of Nodal proteins correlates with clearance.

(A) Upper, representative image and region of interest (red rectangle) for measuring distribution; lower, inset showing magnified region of interest. (B) Normalized distribution profiles and fitting. Error bars indicate standard error of mean (s.e.m). (C) Representative western blots of Nodal proteins harvested from HEK293T cell culture medium at different time points after removal of the source. The Nodal proteins were immuno-precipitated with anti-FLAG antibody and detected by western blot with the same antibody. Schematics on the left show the position of the FLAG epitope tags in each construct. (D) The profile of Nodal protein levels over time after source removal. The data points were fitted with an exponential decay model. Error bars indicate s.e.m.

DOI: http://dx.doi.org/10.7554/eLife.13879.008

Figure 4—figure supplement 1. (Related to Main Figure 4).

Representative immunoblots showing protein degradation over time. Left, representative immunoblots. Equal amounts of sec-GFP-3xFLAG supernatant was added in all of the samples as input controls. Right, quantitation and fitting of protein expression levels. Each experiment was repeated three times, and the band intensity of the Nodal or Lefty proteins was normalized to GFP (input control) and to time 0 hr. The plots were then fitted with exponential decay (equation shown on the upper right corner).

DOI: http://dx.doi.org/10.7554/eLife.13879.010