Journal: Cancer Research

Article Title: GDNF–RET Signaling in ER-Positive Breast Cancers Is a Key Determinant of Response and Resistance to Aromatase Inhibitors

doi: 10.1158/0008-5472.can-12-4265

Figure Lengend Snippet: Figure 1. Differential response to LTED in ERþ breast cancer cells. A, top, qRT-PCR analysis for RET (n ¼ 4), GFRA1 (n ¼ 3), and ESR1 (n ¼ 3) in MCF7, T47D, and ZR75-1 parental cells and their LTED derivatives. Data represent mean SEM. Bottom, immunoblotting of total cell protein extracts. High and low exposures with RET antibody are shown. B, 3 day E2-deprived MCF7 and MCF7-LTED cells were serum-starved overnight and cultured for further 48 hours (top) or 96 hours (bottom) 10 pmol/L E2 100 nmol/L ICI182,780. qRT- PCR analysis for RET (n ¼ 3). Data represent mean SEM. C, 3 day E2-deprived MCF7 and MCF7- LTED cells were serum-starved overnight and stimulated with 20 ng/mL GDNF. D, 3 day E2-deprived MCF7 and MCF7-LTED cells were serum-starved overnight 1 mmol/L ICI182,780 and stimulated with 20 ng/mL GDNF. qRT-PCR analysis for PGR (n ¼ 3), EGR1 (n ¼ 4), TFF1 (n ¼ 3). Data represent mean SEM. Two-way ANOVA, Tukey corrected, , P < 0.05.

Article Snippet: Cell-based assays, siRNA transfection, immunohistochemistry, antibodies, and quantitative real-time PCR (qRT-PCR) analysis were as described previously (11, 13) except the TaqMan probe sets (Applied Biosystem): RET (Hs00240887_m1), GFRA1 (Hs00237133_m1), ESR1 (Hs00174860_m1), PGR (Hs01556702_m1) and the following antibodies (Cell Signaling Technology): phospho-RET-Tyr905 (#3221), phospho-c-Jun (#2361), and c-Jun (#9165).

Techniques: Quantitative RT-PCR, Western Blot, Cell Culture