Tgfbr1 kinase inhibitor

For treatment with LY2157299 (20 μM) and LY364947 (1 μM) (TGFBR1 kinase inhibitor, Selleckchem, Houston, TX), 300,000 H1650-M3 cells were plated in a 6 cm² plate. Inhibitor was added the next day and the mixture was incubated for 3–5 days for LY2157299 and 2–3 days for LY364947. The cells were lysed with TRIzol and processed for RNA preparation. To determine IC50 values for various drugs (17-AAG (this drug is not mentioned in main text), cisplatin, doxorubicin, etoposide, erlotinib, epitaxol and tunicamycin), the cells were plated in 96-well plates at 2,000 cells/well. The next day, individual drugs were added to the wells at the indicated concentrations and the mixture was incubated for 5 days. The plates were then washed once with PBS, fixed with 3.7% formaldehyde and stained with crystal violet. Each stained well was destained in 50–100 µl of 10% acetic acid and the absorbance was read in a spectrophotometer at 590 nm.

For treatment with LY2157299 (20 μM) (TGFBR1 kinase inhibitor, Selleckchem, Houston, TX), 300,000 H1650-M3 cells were plated in a 6 cm² plate. The inhibitor was added the next day, and the mixture was incubated for 3–5 days for LY2157299. The cells were lysed with TRIzol and processed for RNA preparation.