Stat3 and CCAAT/enhancer binding protein beta (C/EBP-beta) regulate Jab1/CSN5 expression in mammary carcinoma cells

Table 1 Sequences of oligonucleotides used in RT-PCR, primer extension, cloning site- directed mutagenesis, EMSA and ChIP

Oligonucleotide	Sequence (5' à 3')	Purpose
Jab1 F296-314	ATCTCAGCATTGGCTCTGC	RT-PCR
Jab1 R1094-1076	TAACCTGAGACATCAATC	RT-PCR
Jab1 R883-864	AGAACTCAACGTATTCACCC	RT-PCR
GAPDH F	CCACCCATGGCAAATTCCATGGCA	RT-PCR
GAPDH R	TCTAGACGGCAGGTCAGGTCCACC	RT-PCR
P1	CGCTCCCGGACGCCGCC	Primer extension
+83R	CTGAAAGCTTCGCTCCCGGACGCCGCC	Cloning
-2006F	CTGACTCGAGGTGGTAGTGAGAGG	Cloning
-2958F	CTGACTCGAGCGCATGGGAACCAA	Cloning
-946F	TGAACTCGAGCCTGCTCCCTGTGTC	Cloning
-658F	CTGACTCGAGCCCACTGCCTCCTCG	Cloning
-472F	CTGAGCTAGCCAACAGACAGCCTT	Cloning
-344F	CTGACTCGAGGAGGCCGAGCCTGC	Cloning
-127F	TGAGCTAGCGTCCCGGAAAGGTCCCC	Cloning
-59F	TGACTCGAGCTGCCCCAAGAGTC	Cloning
-31F	CTGAGCTAGCGTTCCCGTGGTGCGG	Cloning
CEBP-Mut	GCCTTACCTTTTAGTCTTTCctgAAACTTATCTC	Site-directed mutagenesis
GATA1-Mut	CAACAAACTTgagTCATTTAAGGTACCTATACCC	Site-directed mutagenesis
GATA1-Del	GTCTTTCAACAAACTT....CATTTAAGGTACC	Site-directed mutagenesis
-462/-445 CEBP WT	CCTTACCTTTTAGTCTTTCAACAAACT	EMSA
-462/-445 CEBP Mut	CCTTACCTTTTAGTCTTTCctgcAAACT	EMSA
-444/-417 GATA1 WT	AATCATTTATCTCATTTAAGGTACC	EMSA
-444/-417 GATA1 M1	GGAGTCATTTAAGGTACCTATACCC	EMSA
-472F	CTGACTCGAGCAACAGACAGC	ChIP
-343R	CTGAAAGCTTCGCAGGCT	ChIP

Lower-case letters represent mutated oligonucleotides.
ChIP, chromatin immunoprecipitation; Del, deletion; EMSA, electrophoretic mobility shift assay; F, forward; Mut, mutant; M1, mutant-1; P, primer; R, reverse; RT-PCR, reverse-transcription polymerase chain reaction; WT, wild-type.

ISSN: 1465-542X