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. Author manuscript; available in PMC: 2015 Mar 1.
Published in final edited form as: Mol Cancer Res. 2013 Dec 27;12(3):359–369. doi: 10.1158/1541-7786.MCR-13-0526

Figure 5. RBM24 destabilizes p63 transcript.

Figure 5

(A–C) Ectopic expression of RBM24 has no effect on the level of p63 pre-mRNA. HaCaT (A), ME180 (B), and MIA-PaCa2 (C) cells were transiently transfected with a control vector or a vector expressing RBM24 for 48 h. Total RNAs were isolated and subjected to RT-PCR analysis to determine the level of p63 pre-mRNA, RBM24, and actin mRNA. The level of p63 pre-mRNA was normalized to that of actin and arbitrary set as 1.0 in control cells. The relative fold changes were shown below each lane. (D–F) Knockdown of RBM24 has no effect on the level of p63 pre-mRNA. HaCaT (D), ME180 (E) and MIA-PaCa2 (F) cells were transiently transfected with a control or RBM24 siRNA for 72 h. Total RNAs were purified and subjected to RT-PCR analysis to determine the level of RBM24 and actin mRNA as well as p63 pre-mRNA. The level of p63 pre-mRNA or RBM24 mRNA was normalized to that of actin and arbitrary set as 1.0 in control cells. The relative fold changes were shown below each lane. (G) RBM24 shortens the half-life of ΔNp63 transcript. Upper panel: MCF7 cells were uninduced or induced to express RBM24 for 48 h, followed by treatment with 7 μg/ml actinomycin D for various times. Total RNAs were isolated and then subjected to RT-PCR analysis to determine the level of ΔNp63 and actin transcripts. Lower panel: The level of ΔNp63 transcript was normalized to that of actin and plotted along with a time course to calculate the relative half-life of ΔNp63 mRNA. (H) RBM24 shortens the half-life of TAp63 transcript. Upper panel: MIA-PaCa2 cells were transiently transfected with a control vector or RBM24-expressing vector for 48 h, followed by treatment with 7 μg/ml actinomycin D for various times. Total RNAs were isolated and then subjected to RT-PCR analysis to determine the level of TAp63 and actin transcripts. Lower panel: The level of TAp63 transcript was normalized to that of actin and plotted along with a time course to calculate the relative half-life of TAp63 mRNA.