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. 2012 Nov 12;109(48):19691–19696. doi: 10.1073/pnas.1210916109

Fig. 5.

Fig. 5.

Identification of INPP5E interacting proteins and their roles in INPP5E ciliary targeting. (A) Tandem affinity purification (TAP) of INPP5E and PDE6D. Lysates from HEK293T cells stably expressing FLAG- and S-tagged INPP5E (FS-INPP5E) and PDE6D (FS-PDE6D) were subjected to TAP. Parental cells were used as a negative control. Purified proteins were visualized by silver staining. (B) Requirement of CEP164 and PDE6D for INPP5E ciliary targeting. RPE1 cells were transfected with siRNAs as indicated and localization of endogenous INPP5E (green) was evaluated. Yellow arrowheads indicate the location of centrosomes. (C) Quantitation of ciliated cells in B. Results are averages of at least two independent experiments (mean ± SEM). Double asterisks indicate statistically significant decreases compared with control (P < 0.01, one-way ANOVA). (D) Quantitation of INPP5E-positive cilia in B. Asterisk indicates a statistically significant decrease compared with control (P < 0.05, one-way ANOVA). (E) Comparison of INPP5E-associated centrosomes in CEP164 and PCM1 depleted, unciliated cells. Asterisk indicates statistical significance (P < 0.05, Student t test).