Figure 2.

Dsg3 order is not restored following a low-Ca²⁺ pulse. (A) HaCaT cells were transfected with Dsg3-ΔEA-GFP and were not treated (calcium-dependent desmosomes). Cells were subjected to a low-Ca²⁺ pulse including incubation in normal-Ca²⁺ medium, a switch to low-Ca²⁺ medium at t = −10 min, and a return to normal-Ca²⁺ medium at t = 0 min. Cells were imaged with fluorescence polarization microscopy and Dsg3-ΔEA-GFP intensity, and order factor images are shown over the time course. Scale bar = 5 µm; region of interest scale bar = 1 µm. (B) Order factor (mean ± SD) as a function of time (n = 6 cells from three independent experiments). (C) Order factor at −10, 0, and 90 min, where whiskers indicate the range of the data, and the line and box represent the median ± quartile. (D) Quantification of normalized integrated intensity at −10, 0, and 90 min (ns, not significant, P > 0.05; *, P ≤ 0.05; ***, P ≤ 0.001; ****, P ≤ 0.0001; ANOVA).