Fig 2: AP2M1 expression following apoptosis induction.a HCT116 and HCT116 p21-/- cells have been treated with sn38 for 72 h (5 ng/ml) and apoptosis was evaluated by FACS analysis and the detection of subG1 cells (n = 3 ± s.d.). b Phospho-AP2M1 and p21waf1 expressions have been analyzed by western blot in the indicated cells (n = 3). c, d LS174T or HCT116 cells were treated with sn38 in the presence or absence of Akti 1/2 for 72 h (10 µM) and phospho-AP2M1 and p21waf1 expressions were analyzed as described above (n = 3)

Fig 3: AP2M1 regulates CIS escape and is involved in the transmission of soluble signals from senescent cells.a LS174T cells have been transfected with the corresponding siRNAs and protein expression was analyzed by western blot (n = 3). b Following treatment and siRNA transfection, emergence was evaluated after 10 days. Representative images are shown on the left part of the figure and the quantification of emergent clones is presented on the right part (n = 6; **p < 0.01). c Conditioned media (CM) from parental or senescent cells were collected after 24 h of serum starvation. Migration assays were performed using Boyden inserts. RPMI or conditioned media supplemented with 3% FBS were deposited at the bottom of the well, parental LS174T cells at the top. After 72 h, migrating cells were stained with crystal violet (n = 3). d RPMI or conditioned media collected as above supplemented with 3% FBS were deposited at the bottom of the the well. Parental LS174T cells or cells transfected with the indicated siRNAs were deposited at the top to evaluate the sensitivity of cells expressing or not AP2M1 to soluble signals generated from senescent cells. After 72 h, migrating cells were stained with crystal violet (n = 3)