Fig. 4.

ER stress changes plasma membrane dynamics leading to heterogeneous distribution of CD1d in the plasma membrane of THP1 CD1d cells. (A) iNKT cells activation, measured by IFN-γ secretion, induced by wild-type (Left) and PERK KD (Right) THP1 cells pretreated with thapsigargin 0.03 μM and/or pulsed 3 h with a dose curve of αGC. Experiments are representative of n = 2. (B and C) show the normalized frequency distribution of ln (transit time, ms) of CD1d measured from the basal plane of the cells and analyzed as described in the sFCS analysis section. At least 3 independent measurements were taken to report the values obtained from sFCS data. (D) The color-coded topological distribution of CD1d for a 2 × 2-μm region is shown using fire LUT of Fiji for the different treatment conditions. (E) Quantification of heterogeneity in CD1d distribution on the surface of DMSO and thapsigargin-treated THP1-CD1d cells using the spatial autocorrelation function. m, no. of cells; n, no. of curves; v, no. of autocorrelation curves.