Fig. 2: TACC3 correlates with centrosome clustering (CC) in patients with CA and mediates CC in cells with CA. | Cell Death & Differentiation

Fig. 2: TACC3 correlates with centrosome clustering (CC) in patients with CA and mediates CC in cells with CA.

From: Targeting TACC3 represents a novel vulnerability in highly aggressive breast cancers with centrosome amplification

Fig. 2

A Expression of CC score in breast tumors with high CA20 expression from METABRIC with low vs. high TACC3 expression. B CA status (as % of cell population) of breast cancer cell lines determined by centrin-2 (red) and α-tubulin (green) in interphase and mitosis. Scale bar = 10 µm. C Western blot analysis of TACC3 and the mitotic arrest markers, p-H3 (S10) and Cyclin B1 in JIMT-1.sgCtrl vs. sgTACC3 cells. D Multipolar spindle formation in JIMT-1.sgCtrl vs. sgTACC3 cells as shown by α- (spindle, green) and γ- (centrosome, red) tubulin staining. Scale bar = 10 µm. E Quantification of mitotic cells with multipolar spindles from D. F Multipolar spindle formation in BO-264-treated JIMT-1 and MDA-MB-231 (MDA-231, here and for all figures) cells as shown by α- (spindle, green) and γ- (centrosome, red) tubulin staining. Scale bar=10 µm. G Quantification of mitotic cells with multipolar spindles from F. H Western blot analysis of mitosis markers in JIMT-1 cells synchronized at mitosis using nocodazole followed by release into fresh vs. BO-264-containing media. I Dose response curve of MDA-MB-468 (MDA-468, here and for all figures) cells 72 h after treatment with BO-264 upon CA induction with 1 µM of cytochalasin D for 20 h. J IF staining of α- (green) and γ- (red) tubulin in MDA-MB-468 cells treated with 1 µM of cytochalasin D for 20 h followed by 24 h treatment with 5 µM BO-264. Scale bar = 10 µm. K Quantification of mitotic cells with multipolar spindles from J. L BO-264 dose response curve of MDA-MB-468 cells transfected with control vector or PLK4 to induce CA. M IF staining of α- (green) and γ- (red) tubulin in MDA-MB-468 cells transfected with control or PLK4 vector for 24 h followed by treatment with 5 µM BO-264 for an additional 24 h. Scale bar = 10 µm. N Quantification of mitotic cells with multipolar spindles from M. O Correlation of PLK4 expression with CA20 score in breast cancer patients from METABRIC. P Expression of PLK4 in different breast cancer subtypes in METABRIC dataset. Q TACC3 mRNA expression in breast cancer tumors with low vs. high PLK4 expression. R Survival analyses in breast cancer patients based on PLK4 and TACC3 expression. CC centrosome clustering.

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