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. Author manuscript; available in PMC: 2020 Mar 30.
Published in final edited form as: Cell Death Differ. 2019 Oct 1;27(4):1186–1199. doi: 10.1038/s41418-019-0407-5

Figure 2. p27 is induced after PML silencing.

Figure 2

(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 3 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 2 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 3 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.