Fig. 5. Cell surface expression of ∆F508-CFTR is suppressed by knockdown of specific motor proteins.
Immunofluorescent analysis was conducted on ΔF508-CFTR in cells subjected to siRNA treatment targeting KIF1A, KIF5A, and DHC1. Hela cells underwent sequential transfections with the indicated siRNAs (48 h) and plasmids (24 h), followed by immunostaining using anti-CFTR (green, Alexa Fluor 488) and anti-HA (red, Alexa Fluor 568) antibodies. The anti-HA immunofluorescence signal indicates the expression level of ARF1-Q71L. (A) ΔF508-CFTR was predominantly observed in perinuclear regions. (B) Co-transfection with ARF1Q71L in Hela cells resulted in an overall intracellular distribution and cell-surface expression of ΔF508-CFTR. Arrows represent the cell-surface rescue of ΔF508-CFTR by ARF1-Q71L. (C) Treatment with siRNAs against KIF1A (siKIF1A), (D) treatment with siRNAs against KIF5A (siKIF5A), and (E) treatment with siRNAs against DHC1 (siDHC1) abolished the ARF1Q71L-induced cell surface expression of the mutant CFTR. Three independent experiments yielded consistent results. Scale bar: 10 µm. CFTR, cystic fibrosis transmembrane conductance regulator.
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