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. 2015 Nov 15;128(22):4063–4073. doi: 10.1242/jcs.160556

Fig. 3.

Fig. 3.

Effect of depletion of Arf4 and Asap1 on the ciliary targeting of PC1. (A) qRT-PCR analysis of Arf4 and Asap1 depletion by lentiviral shRNA in IMCD-3 cells. The relative knockdown (KD) efficiency was over 99% for Arf4 and ∼75% for Asap1. SCRB, scrambled shRNA control. (B) Depletion of Arf4 and Asap1 in IMCD-3 did not affect ciliogenesis of these cells as evaluated by DAPI and acetylated α-tubulin (ac-α-tub, red) staining. Scale bar: 20 µm. (C) Depletion of Arf4 and Asap1 in IMCD-3 did not affect PC1 trafficking to cilia. PC1 was visualized with an antibody against GFP. Representative images taken for the same exposure time in each cell line are shown. Scale bar: 5 µm. (D) Quantification of the percentage of PC1-positive cilia in cells depleted of Arf4 and Asap1. At least 50 ciliated, PC1 transfected cells were counted for the presence of YFP-PC1 signal on cilia under each condition from at least three independent experiments.