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. Author manuscript; available in PMC: 2016 Sep 1.
Published in final edited form as: Cancer Res. 2015 Jun 30;75(17):3623–3635. doi: 10.1158/0008-5472.CAN-14-2999-T

Figure 6. FGF/RAS-dependent resistance is reversible.

Figure 6

(A) Removal of HRAS(G12V) restores LDE225 sensitivity. SMB(HRAS) cells were infected with lentivirus expressing Flp or tdTomato (control), then assayed by relative survival at indicated LDE225 concentrations (mean ± s.e.m., n = 3).

(B–C) Shh signaling assayed by Immunoblot and qRT-PCR. mean ± s.d., n = 3, ** p < 0.01, *** p < 0.001, one-way ANOVA, Bonferroni correction.

(D–E) SMB21(GF) and SMB55(GF) cells were cultured in media without GF for 3 weeks. (D) Relative survival was then analyzed in indicated LDE225 concentrations (72 hrs). (mean ± s.e.m., n > 9). (E) qRT-PCR analysis of Atoh1, Gli1. mean ± s.d., n = 6, *** p < 0.001, one-way ANOVA with Bonferroni.

(F) Relative survival for SMB21 and SMB21(HRAS) cells treated with MEK inhibitor PD325901 (120 hrs) (mean ± s.e.m., n = 4).

(G) MEK inhibitor PD325901 (0, 10, 100, 1000 nM for 24 hrs) reduce phospho-Erk analyzed by immunoblot.