Two-color labeling tracks changes in relative surface expression.
(A) Representative fluorescence intensity histograms for Brefeldin
A (BFA) treated cells over 18 h. (B) Median fluorescence values for
labeled samples for surface (MG-BTau) and internal (MHN-Ester). The
normalized relative surface expression (MG-Btau/MHN-Ester) is shown
as a measure of surface residence. (C) RSE declines to 77.7% after
6 h, and to 38.4% after 18 h. Error bars are 95% confidence intervals, n = 9–12 replicates per condition from 4 experiments
for all conditions except for 18 h, for which n =
9 from 3 experiments. (D) Representative fluorescence intensity histograms
for dynasore treated cells over 6 h. (E) Quantification of surface
and internal labeling in dynasore treated cells. Median fluorescence
values were averaged across samples. (F) RSE of dynasore treatment
shows a steady increase to 127.0% after 6 h (n =
9 biological replicates from 3 experiments). Error bars are 95% confidence
intervals, one-way ANOVA with Dunnett’s multiple comparison
test. * p ≤ 0.05, ** p ≤
0.01, *** p ≤ 0.001, **** p ≤ 0.0001.