Fig. 2
Mammalian exocyst complex comprises two subcomplexes that can localize to the PM independently. a Quantitative full-scan LC–MS/MS analysis of endogenous EXO70-GFP pulldowns from intact or Sec8-depleted cells. Inputs for the samples were calibrated and normalized using MRM–MS analysis on the same samples. The schematic shows the experimental design in which EXO70-GFP was captured using GFP-Trap beads and SEC8 was depleted by shRNA. b Heatmap summarizing relative binding of SEC6, SEC8, EXO70, and EXO84 to SEC3-GFP in SEC10-depleted cells compared to control shRNA-treated cells, as assessed by western blot. Also see Figure S2H-I. c TIRFM images of EXO70-GFP and SEC3-GFP in control or SEC10 shRNA-treated cells. Scale bars 20 µm. d Quantification of relative fluorescence intensities in C. e Quantification of fluorescence intensities from TIRFM images of SEC5-GFP cells treated with control or SEC10 shRNA. f TIRFM images of double knock-in NMuMG cells, showing localizations of EXO70-GFP and SEC5-Sc at the PM in control, SEC8 shRNA, and SEC3 shRNA-treated cells. Scale bars = 20 µm. g Quantification of relative fluorescence intensities in D. Center lines show the median; box limits indicate 25th and 75th percentiles; whiskers extend 1.5X the IQR from the 25th and 75th percentiles. Experiments were repeated at least three times with similar results
