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. 2015 Nov 4;5:16173. doi: 10.1038/srep16173

Figure 5. Dysregulation of the Cdc25B-Cdk2-NPM axis contributes to centrosome overduplication in Lats1−/− MEFs.

Figure 5

(A) Lats1−/− MEFs transfected with Cdc25B siRNA were immunostained with an anti-γ-tubulin antibody and counterstained with Hoechst 33258 to visualize DNA. GL2 was used as a negative control. The percentages of mononuclear cells with more than two γ-tubulin foci are shown. Data represent mean and standard deviation of three independent experiments (n > 100 cells). (B) Lats1+/+ and Lats1−/− MEFs were treated with or without 180 μM roscovitine (Rosc) for 24 hr. The cells were stained with anti-centrin and anti-γ-tubulin antibodies. The percentages of mononuclear cells with more than two centrosomes are shown (n > 100 cells). (C) Lats1+/+ and Lats1−/− MEFs were immunoblotted with anti-NPM-pT199, anti-NPM, and anti-Lats1 antibodies. α-tubulin was analyzed as the loading control. (D) Lats1−/− MEFs were transfected with 6Myc-Vec, -Lats1-WT, and -Lats1-KD. NPM-pT199 was detected by western blotting. Signal intensity was measured using ImageJ software and normalized to α-tubulin. (E) A model for regulation of centrosome duplication by Lats1.