Table 1.

Experimental condition

Real-time PCR
Ligands	Probes	Conditions
EGF	Hs00153181_M1 (Applied Biosystem)	50°C 2′–95°C 10′–95°C 15′, 60°C 1′ × 40 cycles
TGFα	Hs00608187_M1 (Applied Biosystem)
PTEN	Hs02621230_S1 (Applied Biosystem)
S6	Hs01058685_g1 (Applied Biosystem)
Molecular analysis
Gene	Exon	Sense	Antisense	Annealing temperature (°C)
EGFR	18	ctgaggtgacccttgtctc	tacagcttgcaaggactctg	60 × 40 cycles
	19	agatcactgggcagcatgtg	cagctgccagacatgagaa
	20	cttctggccaccatgcgaa	ccaggcaaactctgctatcc
	21	tcttcccatgatgatctgt	cctggtgtcaggaaaatgct
PI3K	9	gggaaaaatatgacaaagaaagc	ctgagatcagccaaattcagtt	58 × 35 cycles
	20	ctcaatgatgcttggctctg	tggaatccagagtgagctttc
PTEN	5	tgcaacatttctaaagttacctact	gaggaaaggaaaaacatcaaaaa	55 × 40 cycles
	6	tttttcaatttggcttctcttttt	tgttccaatacatggaaggatg
	7	cagttaaaggcatttcctgtg	ttttggatatttctcccaatgaa
	8	tgtcatttcatttctttttcttttc	aagtcaacaacccccacaaa
	9	tcatggtgttttatccctcttg	tgagtcatatttgtgggtttca
K-Ras	1	tggtggagtatttgatagtgt	catgaaaatggtcagaga	56 × 35 cycles
	2	ggtgcactgtaataatccagac	tgatttagtattatttatggc	50 × 45 cycles
B-Raf	11	tccctctcaggcataaggtaa	cgaacactgaatatttcctttgat	55 × 40 cycles
	15	tcataatgcttgctctgatagga	ggccaaaaatttaatcagtgga	56 × 40 cycles
Immunohistochemical analysis
Antibody	Clone	Company	Dilution	Antigen retrieval	Positive control
PDGFRB	sc-339	Santa Cruz Biotecnology	1:100	6′ at 95°C mM citrate buffer, pH 6	Human breast carcinoma
EGFR	K1492	Dako	Procedures performed in accordance with manufacturer's protocol
Her2	Polyclonal	Dako	1:2000	6′ at 95°C, pH 6	Human breast carcinoma

TGFα, transforming growth factor α.