Figure 3

Figure 3

Functional analysis of the Y205F mutant using dye uptake assay and Fluorescence Recovery After Photobleaching (FRAP). (A) Fluorescent images are showing dye uptake of panx1a WT and mutants 10 min after EtBr application. EYFP-transfected cells were used as a negative control. Scale bar: 10µm. (B) Total EtBr fluorescence measured 10 min post dye application shown as a bar graph. Wild type panx1a: n = 17, Y205A: n = 14, Y205F: n = 16, W123A: n = 13, EYFP: n = 13. (C) EtBr fluorescence measured throughout the 10 min with one-minute intervals. (D) FRAP analysis of WT panx1a and Y205F mutant showing selected regions (white rectangles) pre bleaching, immediately after bleaching, and recovery 5 s and 50 s post bleaching. Scale bar: 10um. (E) Total % recovery, 50 s post photobleaching. Wild type panx1a: n = 35, Y205F: n = 33. (F). % recovery measured throughout 50 s with five-second intervals. Error bars show the standard deviation of the mean. ***p < 0.001, N.S., not significant.