Fig. 15.

LATS2-knockout HeLa-S3 cells exhibit multinucleated phenotype, telophase delay, and chromosome missegregation with lagging chromosomes. (A) Exponentially growing LATS2-knockout (LATS2 KO) HeLa-S3 cells and parental HeLa-S3 cells were fixed, and immunostained with anti–α-tubulin antibody (red) and Hoechst 33258 (gray or blue). Scale bar, 10 μm. (B) Percentage of cells with the indicated nuclear number in LATS2 KO (black) and parental (gray) HeLa-S3 cells. Data represent the average of ∼200 cells from three independent experiments. (C) Percentage of mitotic cells (including cells in prometaphase, metaphase, anaphase, and telophase) in asynchronous LATS2 KO and parental HeLa-S3 cells (blue bars). Data represent the average of ∼240 cells from three independent experiments. (D) LATS2 KO and parental HeLa-S3 cells were fixed 1 h after release from nocodazole arrest, immunostained with anti–α-tubulin antibody and Hoechst 33258, and counted by microscopy. Graphs show the percentage of mitotic cells at the indicated stage. Data represent the average of ∼80 mitotic cells from three independent experiments. (E) Percentage of cells with a chromosome bridge or lagging chromosomes during anaphase. LATS2 KO and parental HeLa-S3 cells were synchronized, stained, and counted as in D. Data represent the average of ∼40 anaphase cells from three independent experiments. (F) Percentage of cells undergoing multipolar division during telophase. LATS2 KO and parental HeLa-S3 cells were synchronized, stained, and counted as in D. Data represent the average of ∼30 telophase cells from three independent experiments. (G) Percentage of cells with multipolar spindles during metaphase. LATS2 KO and parental HeLa-S3 cells were synchronized, stained, and counted as in D. Data represent the average of ∼40 metaphase cells from three independent experiments.