CEP131 Abrogates CHK1 Inhibitor-Induced Replication Defects and Is Associated with Unfavorable Outcome in Neuroblastoma

Figure 4

CEP131 promotes cell growth and rescues replication defects in NB-39-nu cells. (a) Western blotting for NB-39-nu cells stably integrated V5-tag-conjugated CEP131 gene (CEP131-V5tag), V5-tag-conjugated LUCZ gene (LUCZ-V5tag), or No lentiviral transduced cells (NB-39-nu). (b) Colocalization of endogenous and exogenous CEP131 was detected by immunofluorescence staining analysis (yellow). The lentiviral-mediated exogenous CEP131 was detected with anti-V5 antibody (green). Both endogenous and exogenous CEP131 proteins were detected with anti-CEP131 specific antibody (red). (c) AlamarBlue cell viability assay. LUCZ-V5tag cells (LUCZ) and CEP131-V5tag cells (CEP131) were treated with 1 μM PF-477736 (_CHK1i) or DMSO (_NT) for 48 h and analyzed for cell viability. Data are presented as the mean ± SD of triplicates. (d) Detection of S phase nuclei. CEP131-V5tag cells and LUC-V5tag cells were treated with 1 μM PF-477736 (CHK1i) or DMSO (NT) for 30 min and then exposed EdU for 10 min. S phase nuclei (red) were scored. . . ns: not significant.

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