Figure 2From: PintlincRNA connects the p53 pathway with epigenetic silencing by the Polycomb repressive complex 2PINT modulates cell proliferation and apoptosis. (A) Inhibition of Pint. Pint levels were detected by quantitative real time (RT-qPCR) in p53-restored doxorubicin-treated p53LSL/LSL MEFs 36 hours after transfection with two Pint-specific anti-sense oligonucleotides (ASOs) (ASO1 and ASO2), two control ASOs (control ASO -1 and -2), or a blank (PBS) control, and 12 hours of doxorubicin treatment. Values normalized to Gapdh and are the mean ± SD of three replicates. (B) Pint positively regulates cell proliferation. Relative number of p53-restored p53LSL/LSL mouse embryonic fibroblasts (MEFs) transfected with ASOs for Pint inhibition, and treated with doxorubicin from 24 h post-transfection. Cell numbers are determined by MTS assay. Values are mean ± SD of three replicates. (C) Overexpression of Pint. Pint levels where measured like in (A) in p53-restored doxorubicin-treated p53LSL/LSL MEFs 36 hours after transfection and 12 hours of doxorubicin treatment with Pint A isoform expressing plasmid or an empty plasmid as control. (D) Pint positively regulates cell proliferation. Cells were transfected as in (C) and treated with doxorubicin from 24 hours post-transfection. (E,F). Negative effect of Pint on apoptosis induction. Apoptosis levels were determined by quantification of caspase 3/7 levels after (E) inhibition or (F) overexpression of Pint in p53-restored p53LSL/LSL MEFs treated with doxorubicin. Values are the mean ± SD of three replicates. (G,H). Effect of Pint on cell cycle regulation. Relative cell numbers in each cell cycle phase were determined by fluorescence-activated cell sorting (FACS) of bromodeoxyuridine (BrdU) incorporation and propidium iodide (PI) staining of p53-restored p53LSL/LSL MEFs treated as in (A) or (C). Percentages of cells in each phase are represented and values are the mean ± SD of three replicates.Back to article page