Figure 4.

Microtubule organization and ciliogenesis require CEP290 function. (A) The depletion of CEP290 from hTERT-RPE cells causes a collapse of microtubule networks, which is accompanied by an apparent decrease in cell size. (B–C) hTERT-RPE cells were shifted from 10% serum to serum-free medium 24 h after transfection with the indicated siRNAs and cultured for additional 60 h before fixation. Ciliated cells were identified by anti-Ac-tub antibody staining. Basal body was labeled by anti-γ-tub antibody. The depletion of either CEP290 or PCM-1 inhibits ciliogenesis. (C) Histogram quantifying the data shown in (B). More than 200 cells from two independent experiments were counted for each bar. Error bars represent SD. (D) Representative images of hTERT-RPE cells stained with the indicated antibodies after CEP290 siRNA transfection and serum starvation. Both ninein and pericentrin staining of CEP290-depleted cells were indistinguishable from those of control cells. Scale bars: (A) 10 µm; (B and D) 20 µm.