Fig. 3: Flow cytometry analysis of GreenT-EC cell surface exposure.

Twelve combinations of export signal peptides and transmembrane/anchoring domains were used to target GreenT-EC to the surface of the cell. HeLa cells were transiently transfected with each construct and analyzed using flow cytometry. a Density plots of the negative control (non-transfected) indicating the gating strategy for selecting GreenT-EC positive single cells (indicated in slashed lines). Both the negative and the rest of the constructs displayed were resuspended in a MOPS buffer containing 3 mM CaCl₂. b The fluorescence of GreenT-EC positive gated cells was further analyzed before and after the addition of EGTA (final concentration of 7.5 mM). Box-plots 90-10% are presented (whiskers are 90 and 10 percentiles, boxes are 75 and 25 percentiles and the center line correspond to the median). c Summary table compiling the main parameters of the experiment: Number of cells gated as GreenT-EC positive in each condition (n=cell count), mean fluorescence of GreenT-EC (mean fluoresce.), standard error of the mean (s.e.m) and the response calculated between the mean GreenT-EC fluorescence values before and after the addition of EGTA (ΔF/F₀). Source data are provided in the Source Data file.