Research Paper Volume 1, Issue 1 pp 58—67

Figure 4. ΔNp63α increases levels of the mTERT-spliced isoforms via protein interaction with ABBP1. Mouse epidermal keratinocytes (2x10⁶ cells) expressing heterozygous p63-/+ and ΔNp63α transgene. (A) Cells were tested for the levels of ΔNp63αand ABBP1 by immunoblotting and ABBP1ΔNp63αprotein complexes using immunoprecipitation (IP) with an antibody to ABBP1 followed by immunoblotting with an antibody to ΔNp63α. As a control, the protein level of β-actin was monitored. (B) Cells were examined for the expression of the mTERT and mTR transcripts using RT-PCR. GAPDH was used in RT-PCR assay, as a control. (C) The relative expression of TERT and TR was quantitatively analyzed and plotted as bars using the Microsoft Excel software. All of the data (mean +SD) were from at least three independent experiments. Samples: cells from p63-/+ mice, 1- TERT/GAPDH ratio; 2- TR/GAPDH ratio; cells from the ΔNp63αtransgenic mice, 3- TERT/GAPDH ratio; 4- TR/GAPDH ratio. PCR experiments with the 2164/ 2620 set of primers generated three products that represent the full-length TERT transcript (457 bp), the α-splice transcript (421 bp), and the β-splice transcript (275 bp). Sequence analysis revealed that the longer transcripts were full-length one and the shorter transcripts were α and β- spliced messages of mTERT.