Table 1 Oligonucleotide sequences used for RT-PCR, RT-qPCR analyses and morpholino design
Primers for ΔNp53 PCR amplification (see Fig. 1A) | ||
|---|---|---|
Gene | Forward | Reverse |
E2/E11 | GCAAAACGACAGCCAAGAGT | ACGTCCACCACCATTTGAAC |
I2/E5 | CAGCCATGTCAGGTTGCTAT | CACCTTAATCAGAGTCGCTTC |
Primers for quantitative PCR (see Fig. 5) | ||
Gene | Forward | Reverse |
p21 | GGAAAACATCCCGAAAACACC | TGTGATGTTGGTCTGTTTGCG |
ΔNp53 | CCTGATACACAACGCTCCTCT | TGATGTCCCAGCAAGAGCC |
Δ113p53 | ATTCTGTGTGACATTACAAGACCAGG | GTTTATTCAGGTCCGGTGAATACC |
ELF1A1 | CTTCTCAGGCTGACTGTGC | CCGCTAGCATTACCCTCC |
Primers for semiquantitative PCR (see Figs. 1C/2A) | ||
Gene | Forward | Reverse |
Δ113p53 | TTTGGAGGGAGATGTTGGTC | GTTATCTCCATCCGGGGTTC |
ΔNp53 | TGCATCTGTCGAACGTATTG | GTTATCTCCATCCGGGGTTC |
β-Actin | GATGTGGATCAGCAAGCAGGAGTA | CGAGAGTTTAGGTTGGTCGTTCGT |
Morpholino sequences | ||
Gene | Sequence | |
p53 | 5'GCGCCATTGCTTTGCAAGAATTG3' | |
ΔNp53 | 5'AGGTACATTATAGCAACCTGACATG3' | |
p21 | 5'TAATAAAGAGGTCTGACCTGTGATG3' | |
standard control | 5'CCTCTTACCTCAGTTACAATTTATA3' | |
ΔNp53 mismatch control | 5'AGcTAgATTATAGgAACCTcACtTG3' | |