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. 2019 Sep 16;17(1):33–46. doi: 10.1080/15476286.2019.1662268

Figure 2.

Figure 2.

P27(3ʹUTR) reporter mRNA recapitulates post-transcriptional regulation in CP-treated cells. (a) Schematic representation of GFP-T and GFP-T-p27(3ʹUTR) reporter constructs. GFP-T cells lack exogenous p27 3ʹUTR sequences. (b) Changes in GFP and endogenous p27 mRNAs in CP-treated (+CP) compared to untreated (-CP) GFP-T and GFP-T-p27(3ʹUTR) inducible cell lines. Transcript levels were quantified with RT-qPCR and normalized to β-actin mRNA. Error bars represent SEM, n = 3. *P < 0.05, **P < 0.01. (c) mRNA decay of GFP-T-p27(3ʹUTR) in untreated (-CP) and CP-treated (20 µM CP for 15 h) cells. The half-life of GFP and c-myc mRNAs was determined relative to β-actin with RT-qPCR and plotted as the mean, considering ‘one phase decay equation’ implemented in GraphPad Prism. Error bars represent SEM, n = 2.