A. Sensitivity of MDA-MB-134 cell lines to FGFR1 siRNA (siFGFR1). Cells were transfected with siFGFR1, or siCON non-targeting control, and survival assessed six days later with Cell Titre-Glo® cell viability assay (Promega). MDA-MB-134 obtained directly from MD Anderson were sensitive to FGFR1 knockdown (p<0.001, Student's T Test), but not MDA-MB-134 obtained from ATCC. Error bars SEM of three repeat experiments.
B. Left: Transfection of FGFR1 amplified cell lines with siCON or siFGFR1, and siPLK1 as a positive toxicity control, with survival assessed at 5-7 days transfection. Right: FGFR1 expression by quantitative RT-PCR in SUM44 cells transfected with siFGFR1, or siCON, 72 hours prior to RNA extraction.
C. FGFR1 amplified cell lines were grown for 96 hrs in media supplemented with a range of concentrations of PD173074 pan FGFR tyrosine kinase inhibitor, and survival expressed relative to that of untreated cells. The SUM52PE breast cancer cell line that harbours FGFR2 amplification, and is highly sensitive to FGFR inhibitors, was used a positive control (26).
D. CAL120 cells were grown in soft agar with or without continuous exposure to 1μM PD173074. Example micrographs at 4X power from wells with and without PD173074. Bar chart: mean colonies per well from 3 repeats (without 25.3 colonies vs with PD173074 0 colonies, p=0.008, Student's T test).