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Figure 2 | BMC Molecular Biology

Figure 2

From: Cdc5 influences phosphorylation of Net1 and disassembly of the RENT complex

Figure 2

Overexpression of Cdc5(DB▲) triggers Cdc14 release. (A) Overexpression of Cdc5(DB▲) promotes release of Cdc14 from the nucleolus even in cells with short spindles. CDC14-MYC9 cells with (WY333) or without (WY201) integrated GAL-CDC5(DB▲) were grown in YP + 2% raffinose (GAL promoter uninduced), arrested in G1 by α-factor, supplemented with galactose to 2% for 0.5 hour (GAL promoter induced), and released into YP + 2% galactose at time = 0. At indicated time points after release, samples were fixed, and subjected to indirect immunofluorescence with anti-tubulin and 9E10 antibodies to visualize the mitotic spindles and Cdc14-Myc9, respectively. The percentages of cells with short spindles and diffused Cdc14 were calculated. (B) Overexpression of Cdc5(DB▲) triggers Cdc14 release in cells arrested in mitosis. NET1-MYC9 cells with (WY331) or without (WY53) integrated GAL-CDC5(DB▲) were grown to exponential in YP + 2% raffinose, arrested in mitosis by nocodazole (10 μg/ml final), and supplemented with galactose to 2% for four hours. Samples were subjected to indirect immunofluorescence with anti-Cdc14 and 9E10 (to detect Net1-Myc9).

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