1. Fig. 3. Differentiation of AZD4785 from MAPK pathway inhibitors in vitro.
Differentiation of AZD4785 from MAPK pathway inhibitors in vitro. (A) Western blot analysis of MAPK and PI3K signaling in NCI-H358 and PC9 cells treated with AZD4785 and CTRL ASO for 72 hours or with selumetinib, SCH772984, or DMSO for 24 hours. (B) Expression of KRAS, DUSP6, and ETV4 mRNA measured by qRT-PCR in NCI-H358 and PC9 cells treated with a dose range of AZD4785 for 72 hours or with selumetinib or SCH for 24 hours. Expression was normalized to GAPDH and expressed relative to PBS control. Data are from a representative experiment (n = 2) showing mean expression of technical replicates and SD. (C) Western analysis of MAPK pathway signaling in NCI-H358 and PC9 cells after monotherapy or combination treatment with AZD4785, CTRL ASO, and selumetinib. ASO treatment was done for 72 hours, and selumetinib treatment was done for 24 hours. For combination treatments, selumetinib was added 48 hours after dosing with AZD4785 for the final 24 hours of incubation. (D) NCI-H358 and PC9 cells grown in soft agar were treated with selumetinib in combination with CTRL ASO or AZD4785. Data are from a representative experiment from a minimum of two showing mean colony number and SD.
Sarah J. Ross et al., Sci Transl Med 2017;9:eaal5253
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