Extended Data Fig. 7: CDK4/6i-resistant cells express CDK6 as a thermostable, weak HSP90 client protein.
From: Distinct CDK6 complexes determine tumor cell response to CDK4/6 inhibitors and degraders
a, Comparison of total peptide-spectrum match (PSM) for CDK6-interacting proteins by mass spectrometry in KMS-12-PE and Calu6. b, Cell lysates from Colo205 and Calu6 were either subjected to Co-IP with a CDK6 antibody followed by immunoblotting with HSP90, CDC37 and CDK6, or immunoblotted with the indicated antibodies. c, The indicated cell lines were treated with increasing concentrations of Ganetespib (GAN) for 24 hr. Lysates were subjected to immunoblotting with the indicated antibodies. d, The indicated cell lines were treated with 40 nM Luminespib (LUM) at the indicated time points. Lysates were subjected to immunoblotting with the indicated antibodies. e. Calu6 cell line expressing Dox-inducible shCDC37 were treated with or without 0.1 µg/ml doxycycline for 72 hr and cell lysates were subjected to immunoblotting with the indicated antibodies. f, CDK4-dependent cell lines were treated with 30 nM GAN for the indicated time points. Lysates were subjected to immunoblotting with the indicated antibodies. g, KMS-12-PE and Calu6 were treated with increasing concentrations of GAN for 24 hr. Lysates were subjected to immunoblotting with the indicated antibodies. h, Lysates from A375 cells ectopically expressing V5-CDK6 or V5-CDK6 S178p were immunoprecipited with a V5 antibody. The immunoprecipitates were subjected to kinase assay with recombinant Rb protein as substrate.
