Fig. 4: Minute-resolution temporal profiling of the STS1 interactome.

a Workflow of proximity proteomic analysis with five EGF stimulation time-course. HeLa cell line with stably expressed APEX2-FLAG-STS1 were stimulated with EGF and labeled with BP5 as indicated. b Volcano plots of EGF stimulation-dependent STS1 interactome at each stimulation point. Significantly changed proteins are marked in red (n = 3 independent biological experiments). HeLa cells without EGF stimulation was used as common control. c. The temporal profiles of the STS1 interactome in living cells. The size of each dot is proportional to the relative abundance of the 35 interacting proteins. The CV (coefficient of variation) distribution is color-coded with different intensities. d. Four manually curated clusters of the STS1 temporal interactome according to curve similarity. The numbers and red curves indicate the proteins in each cluster and the average values of all the proteins, respectively. e, f Validation of STS1 interaction with GRB2 by streptavidin pull-down of APEX2-FLAG-STS1 and western blots (n = 3 independent biological experiments). Data are presented as mean values ± s.d. (error bars). *p-value is 0.0359. **p-value < 0.0001. n.s. not significant (according to two-sided Student’s t tests with adjustments made for multiple comparisons). g, h Validation of STS1 interacts with GAB2 by streptavidin pull-down of APEX2-FLAG-GAB2 and western blots (n = 3 independent biological experiments). Data are presented as mean values ± s.d. (error bars). Quantification of the WBs in f and h were presented in Supplementary Fig. 12b, c. Source data are provided as a Source Data file.