Figure 7.

Colocalization of PCM1 with subcellular CROCC. (A) The CEP250 and CROCC KO RPE1 cells were treated with 20 μM nocodazole for 2 h and subjected to coimmunostaining analysis with antibodies specific to PCM1 (cyan) and centrin-2 (magenta). (B) Intensities of PCM1 at the centrosome were determined. More than 30 cells per group were counted in three independent experiments. Within each box, the black center line represents the median value, the black box contains the interquartile range, and the black whiskers extend to the 10th and 90th percentiles. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test (***, P < 0.001; n.s., not significant). (C) The cells expressing the ectopic FLAG-CROCC protein were treated with 20 μM nocodazole for 2 h and subjected to immunoprecipitation analysis with the FLAG antibody, followed by immunoblot analyses with antibodies specific to FLAG and PCM1. (D) FLAG-CROCC^FL, FLAG-CROCC^303–1741, and FLAG-CROCC^ΔR3 were stably expressed in the CROCC KO RPE1 cells. The cells were treated with 20 μM nocodazole for 2 h and coimmunostained with antibodies specific to FLAG (cyan) and PCM1 (magenta). (A and D) Scale bars, 10 μm; inset scale bar, 2 μm. Source data are available for this figure: SourceData F7.