Journal: Oncogene

Article Title: Osteopontin-induced migration of human mammary epithelial cells involves activation of EGF receptor and multiple signal transduction pathways.

doi: 10.1038/sj.onc.1206209

Figure Lengend Snippet: Figure 2 Time course showing effect of OPN on expression of TGFa, EGF, and EGFR mRNA by 21PT (PT), 21NT (NT), and MDA-MB-435 (MDA) cells. Near confluent (85–90%) cell cultures were incubated in serum-free medium with 50 mg/ml OPN for 0, 5, 18, or 24 h. Total RNA (10 mg/lane) was analysed by Northern blotting for expression of TGFa (4.8 kb), EGF (5.0 kb) or EGFR (5.6 kb). RNA loading and integrity were verified by assessment of 18S rRNA (2.1 kb). Level of TGFa, EGF, and EGFR are shown in relation to 18S rRNA, expressed as the ratio of densitometry values for the respective bands, plus or minus s.d. on triplicate measurements (TGFa/18S, EGF/18S, EGFR/18S)

Article Snippet: Probes used included TGFa (40 base antisense oligonucleotide derived from sequences encoding the mature human TGFa polypeptide, Derynck et al. (1984), Cedarlane), EGF (0.555 kb EcoRI fragment of human EGF cDNA plasmid phEGF121, containing the 50 untranslated region and encoding amino acids 1–38 of the EGF precursor, Murray et al., 1986, ATCC), and EGFR (1.65 kb ClaI-EcoRI fragment of human EGF receptor cDNA, Xu et al., 1984).

Techniques: Expressing, Incubation, Northern Blot