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. 2006 Oct 1;20(19):2687–2700. doi: 10.1101/gad.1447006

Figure 5.

Figure 5.

Lats2 is required for optimal induction of p53, p21, and Lats2 itself in response to nocodazole. (A) Optimal induction of p53 protein by nocodazole is compromised by Lats2 knockdown. Stably transfected U2OS/siControl and U2OS/siLats2-581 cells were treated with 0.2 μg/mL nocodazole (noc) and harvested at the indicated times. Endogenous p53 was detected using DO1 + PAb18O1 antibodies, and GAPDH was used as a loading control. Quantification of relative p53 levels is shown below each lane. (B) Full induction of Lats2 by nocodazole is dependent on p53 and Lats2 itself. Stably transfected U2OS cells subjected to the indicated shRNA knockdown were treated with 0.2 μg/mL nocodazole (noc) and harvested after 8 h. Endogenous Lats2 was visualized using LA-2 antibodies, and GAPDH was used as a loading control. (C) Basal Lats2 is required for efficient induction of subsequent Lats2 mRNA by nocodazole (positive feedback). Stably transfected U2OS/siControl and U2OS/siLats2 cells were treated with 0.2 μg/mL nocodazole (noc) and harvested at the indicated times. RNA was analyzed by quantitative RT–PCR, as in Figure 3D. “Relative Lats2 mRNA” represents HPRT-normalized RNA levels compared with t = 0 levels in siControl cells.