Figure 4. Expression and localization of CEP290.

(A) Normalized Cep290 gene expression levels were measured by RT-qPCR of WT cells synchronized by serum starvation (G₁), thymidine (S), nocodazole (G₂) and release for 5 hours of these synchronizations, and unsynchronized, APH (400 nM) and ADR (1 μM) (24 hours) treated cells (n = 4; 1-way ANOVA, **P < 0.01). (B) Concomitantly, protein expression was detected by Western blot, using β-actin as loading control. (C) Western blot of chromatin fractionation of WT (–) and Cep290^LacZ/LacZ (+) cells. CEP290 is expressed in the cytosol (GAPDH, tubulin), nucleus (PCNA), and chromatin-enriched fraction (H2AX). Samples were run on parallel gels contemporaneously. (D) Immunofluorescence staining of CEP290 expression (white) of WT cells treated with DMSO, APH (400 nM), and ADR (1 μM) (18 hours). ADR exposure caused increased CEP290 levels. Scale bar: 10 μm.