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. 2017 Jan 9;7:39887. doi: 10.1038/srep39887

Figure 3. Relocalization of Sec16A in Arf1-Q71L-induced unconventional secretion of ΔF508-CFTR.

Figure 3

(a,b) Intracellular localization of Sec16A was analyzed using immunocytochemistry. HeLa cells were transfected with plasmids expressing extracellular loop HA-tagged ΔF508-CFTR with or without Arf1-Q71L. CFTR at the cell surface was immunostained with anti-HA antibodies before membrane permeabilization (green). Then, the total CFTR was stained with anti-M3A7 CFTR antibodies (grey), and Sec16A was stained with anti-KIAA0310 Sec16A antibody (red). In control cells (a), Sec16A puncta were mostly localized in the juxtanuclear area (arrowhead). (b) ER-to-Golgi blockade by Arf1-Q71L evoked the cell-surface expression of ΔF508-CFTR (green) and the redistribution of Sec16A to entire cellular area (arrows, red). (c,d) Sec16A redistribution in cells coexpressing HA-Arf1-Q71L (white dotted line) was compared with that in cells not expressing HA-Arf1-Q71L. Representative immunofluorescence images are shown (c), and the results of multiple experiments (n = 5, each comprising analyses of 5–10 cells, **P < 0.01) are summarized (d). Scale bar: 5 μm, **P < 0.01.