Interaction with CEP90 is critical for centrosomal localization of PCM-1.

<p>(A–C) The CEP90-depleted RPE-1 cells were rescued with Myc-GFP or the RNAi-resistant CEP90 constructs (Myc-CEP90^1–757 and Myc-CEP90^Δ271–363). (A) Forty-eight hours later, the cells were subjected to immunoblot with the antibodies specific to CEP90 and α-tubulin. Asterisks represent non-specific bands. (B) The cells were co-immunostained with the antibodies specific to Myc and PCM-1. (C) The centrosomal localization of PCM-1 was analyzed in over 200 cells per experimental group. The experiments were repeated 4 times. n.s. indicates not significant. (D–E) The CEP90-depleted RPE-1 cells were rescued with RNAi-resistant CEP90 containing a PACT domain (Flag-CEP90r^1–757-PACT and Flag-CEP90r^Δ271–363-PACT). (D) Forty-eight hours later, the cells were co-immunostained with antibodies specific to Flag and PCM-1. (E) The relative intensities of PCM-1 to CEP90-PACT at the centrosome were determined in 20 cells per experimental group. The graphs show the mean values and standard errors (C) or standard deviations (E). Scale bar, 10 μm.