Extended Data Fig. 2: ALK inhibitor-resistant cells exhibit stable resistance in vivo and no longer rely on ALK signalling.
From: BORIS promotes chromatin regulatory interactions in treatment-resistant cancer cells
a, Left, tumour volumes of sensitive and resistant cell xenografts in untreated NU/NU (Crl:NU-Foxn1nu) mice established by subcutaneous injection of 2 × 106 cells into both flanks. Animals were euthanized when tumours reached 1,500–2,000 mm3. Data are mean ± s.e.m., n = 4 per arm. Right, immunoblot analysis of total and phosphorylated ALK in TAE-resistant xenograft tumours (1 and 2) and sensitive and resistant cells in culture. b, Dose–response curves for TAE684 in sensitive and resistant cell lines established from the same tumour xenografts as in a (IC50 values: sensitive, 7.9 nM; resistant, 878.6 nM). Data are mean ± s.d., n = 3 biological replicates. c, Tumour volumes (left) and Kaplan–Meier survival curves (right) of resistant cell xenografts in NU/NU (CrTac:NCr-Foxn1nu) mice treated with TAE684 (10 mg kg−1 by oral gavage once daily) or vehicle control for up to 56 days. Data are mean ± s.e.m., n = 8 per arm. P values determined by Mann–Whitney U test for tumour volumes (P = 0.8404) and by log-rank test for Kaplan–Meier survival analysis (P = 0.8076), both two-sided. d, Dose–response curves for TAE684-sensitive and -resistant cells treated with ceritinib (IC50 values: sensitive, 33.8 nM; resistant, 446.5 nM) or lorlatinib (IC50 values: sensitive, 47.5 nM; resistant, 2,318 nM). Data are mean ± s.d., n = 3 biological replicates. e, Immunoblot analysis of the indicated proteins in sensitive and resistant cells treated with DMSO or 1 μM TAE684 for 6 or 24 h. f, Electropherograms of ALK kinase domain sequencing in sensitive and resistant cells. Arrows show the F1174L mutation characteristic of Kelly cells. HEK293T cells were used as a control for sequencing wild-type ALK. g, Phosphoproteomic analysis of a panel of receptor tyrosine kinases (RTKs) in sensitive and resistant cells. Each RTK is shown in duplicate and the pairs in the corners of each array are positive controls. Numbered RTKs with corresponding names listed on the right represent the highest-phosphorylated proteins. ALK is depicted in red. h, Quantitative reverse transcription PCR (qRT–PCR) and immunoblot analysis of ABCB1 and ABCG2 multidrug transporter expression in sensitive and resistant cells. The qRT–PCR data are means of n = 2 biological replicates. In a (immunoblot), d, f and g, data are representative of two independent experiments (see Supplementary Note 1 for details; for gel source data, see Supplementary Fig. 1).
