Figure 5.
Centrosome Amplification Induces an Early Stress Response that Leads to Altered Secretion
(A) Left, quantification of Ki67 positive cells. Right, cells stained for Ki67. Scale bar: 40 μM.
(B) Left, cells stained for β-galactosidase (blue). Right, quantification of β-galactosidase positive cells after 6 days. Scale bar: 40 μM.
(C) Relative IL-8 secretion (fold, ng/cell) in cells with extra centrosomes (Left) or treated with DoxoR (Right).
(D) Quantification of invasive structures.
(E) Left, cells stained for β-galactosidase (blue). Right, quantification of senescence in RPE-1.PLK4 cells. Note that senescence was assessed by enlarged morphology (purple arrowheads). Scale bar: 40 μM.
(F) Left, quantification of γH2AX foci. Right, cells were stained for γH2AX. L, large nuclei. Number of cells MCF10A.PLK4 −DOX = 469; +DOX = 466; and RPE-1.PLK4 −DOX = 155; +DOX = 115; +DoxoR = 84. Scale bar: 20 μM.
(G and H) (G) Fold change of secreted SASP components in MCF10A and (H) RPE-1 cells.
(I) HMGB1 secretion after 48 hr.
(J) IL-8 secretion after p53 depletion (48 hr).
(K) Quantification of invasive structures.
Graphic in (G) represents 4 independent experiments; for all other graphics, error bars represent mean ± SD from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; ns, not significant.