p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary Cells (original) (raw)
Figure 5
Supernumerary Centrosomes Produce Multipolar Spindles and Correlate with Aneuploidy
(A) BrdU incorporation of HMECs (top) and vHMECs (bottom). Cell cycle analysis were performed using BrdU incorporation for cells that were untreated (−HU), treated for 48 h with HU (+HU), or treated for 48 h with HU, followed by release from HU treatment for 7 to 8 d (+HU → release). DNA replication resumes following release from 48 h of HU treatment.
(B) Microtubule nucleation sites were determined by immunocytochemistry with an antibody recognizing γ-tubulin.
(C) Analysis of HMECs (black: RM9 and RM26 [less than 4 PD]) and vHMECs (red: RM15 [25 PD], RM16 [13 PD]) for multipolar mitosis in untreated (−HU) or exposed to HU followed by release from HU treatment (+HU → release).
(D) Analysis of HMECs (black: RM9 and RM15 [less than 5 PD]) and vHMECs (red: RM9 [21 PD], RM16 [17 PD]), HMECs infected with p16INK4a shRNA (gray) for genomic abnormalities in untreated (−HU) or exposed to HU followed by release from HU treatment (+HU → release). Types of chromosomal abnormalities represented include aneuploidy, structural abnormalities, and telomeric associations. Analysis included at least 100 metaphases. *Statistical significance (p < 0.005) based on comparison of −HU and +HU → release experiments.