Fig. 2

p27 is induced after PML silencing. a Effect of doxycycline-inducible (150 ng ml⁻¹; 3 + 3 days) PML silencing (sh4, sh5) on p27 and PML protein expression (representative of at least three experiments) on MDA-MB-231 cells. b, c Quantification of p27 and PML protein levels along 6 days of doxycycline-inducible PML silencing on MDA-MB-231 cells (n = 3) with two different shRNAs. d Immunofluorescence quantification of nuclear p27 positive cells upon PML inducible silencing on MDA-MB-231 cells (n = 4). e–g Immunofluorescence quantification of the correlation of p27 positive cells and PML levels in these cells (e–f) and representative images of p27 and PML staining (g) upon doxycycline-inducible PML silencing in MDA-MB-231 cells (n = 4). h Effect of doxycycline-inducible (150 ng ml⁻¹; 3 + 3 days) PML silencing (sh4) on RB phosphorylation (Ser780) (representative of three experiments) on MDA-MB-231 cells. i Expression of p27-related cell cycle genes upon PML inducible silencing in MDA-MB-231 cells (n = 3). j Impact of doxycycline-inducible PML silencing (sh4) on p27 and PML protein expression on established MDA-MB-231 xenografts. Error bars represent s.e.m. p, p-value (*^/$p < 0.05, **^/$$p < 0.01, ***^/$$$p < 0.001). One-tailed one-sample t-test (b, c, i) and one-tailed Student’s t-test were used for cell line data analysis (d–f). sh4 and sh5: shRNA against PML, dox: doxycycline. ^#Unspecific band. Molecular weight markers (kDa) are shown to the right