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. 2013 Sep 21;105(19):1485–1495. doi: 10.1093/jnci/djt210

Figure 4.

Figure 4.

VS-6063 downregulated YB-1 phosphorylation and nuclear translocation in taxane-resistant cells by an AKT-dependent pathway. A) Western blot analysis of lysates collected at 0, 5, 10, 30, and 60 minutes after treatment of SKOV3ip-TR cells with VS-6063. The bar graphs at the right show densitometry results. Results were confirmed with triplicate experiments. Error bars indicate standard deviation. Statistical tests were two-sided, and P values were evaluated using Student t test. *P <.001 compared with VS-6063 untreated group (time 0); **P < .01 compared with VS-6063 untreated group. B) Cytoplasmic and nuclear extracts of SKOV3-TR cells were prepared after treatment with AKT1/2 kinase inhibitor (AKTi, 10 µM) for 24 hours, and anti-pYB-1 (Ser 102) and anti-pAKT immunoblots were performed. β-Actin and lamin B were used as loading controls. C) Immunofluorescence staining of YB-1 in SKOV3-TR cells. Cells were treated with or without 10 µM AKT 1/2 kinase inhibitor (AKTi) for 24 hours and then fixed, permeabilized, and incubated at 4° C with the primary pYB-1 (Ser 102) antibody and then with the Alexa 594–labeled secondary antibody.