RhoA Regulation of Cardiomyocyte Differentiation

Table 1

Primers used for PCR.

PCR product	Forward primer (5′-3′)	Reverse primer (5′-3′)	Product size (bp)
Elucidation of chick RhoA gene structure
Chick RhoA Intron1	GCCGGAATTCATGGCAGCCATTCGAAA^1	CTATCAGGACTATCGATTG	282 bp^4
Chick RhoA Intron2	GTGGAGTTGGCTTTGTGGGATAC	GCTTCATTTTGGCCAGCTCT	251 bp^4
Chick RhoA Intron3	AGTGGACCCCGGAAGTGA	TAAGAGAAGGCACCCGGAC	287 bp^4
Amplification of putative mouse RhoA promoter
PromoterLong	CCCGCGGATCCGTGGCGGGCAAAGCTTGCAG^2	CCCGCGGATCCGTCCGGCCTCTTCGCGCT	637
PromoterShort	CCCGCGGATCCAGCAATCGTGGCTGAACTGAG^2	CCCGCGGATCCGTCCGGCCTCTTCGCGCT	264
Generation of dominant mutant of mouse RhoA
mRhoA; N19	CGCCGCTCGAGATGGCTGCCATCAGGAAGAAACTGGTGATTGTT; GGTGATGGAGCTTGTGGTAAGAATTGCTTGCTCATA^3	ACGCGTCGACTCACAAGATGAGGCACCC	582
Primers used for real-time PCR
GAPDH	TCCTACCCCCAATGTGTCCGTC	GCCCAAGATGCCCTTCAGTG	121
RhoA ORF	ATTGATGTGTTTTTCCATTG	CTCCCGTCTCGTGTGCTCGTCATT	151
RhoA 3′UTR	GCTACCAGTATTTAGAAGCCAACCAC	GCTGTTAGAGCAGTGTCAGAAGGAC	88
SRF	TGCCTCAACTCGCCAGACTCTC	TTCAGTGTGTCCTTGGTTTCCC	140
Cardiac 𝛼-actin	GCCAACCGTGAGAAGATGACC	CGCCAGAATCCAGAACAATGC	130
GATA 4	ATGCCGAGGGTGAGCCTGTATG	CTTCCGTTTTCTGGTTTGAATCC	110

1Incorporated EcoR1 and 2BamH1 sites are underlined.
3ATG translation start codon and introduced N19 mutation are shown in bold.
4Expected size in absence of intron.