Fig. 5
In vitro therapeutic efficacy and in vivo pharmacodynamics. a Western blotting showing the levels of each of the 6 kinases in the signature, in addition to their non-phosphorylated controls where applicable, in 10 human TNBC cell lines, the transplantable murine TNBC tumor model 4T1, and 2 patient-derived xenografts (PDXs). The three targets against which clinical-grade drugs are available are highlighted in red. b Colony assays (MDA-MB-231) showing the differences between single-target versus two-target pharmacological blockade. For each of the 15 possible 2-by-2 drug combinations using the 6 agents against the kinases in K-high, a representative vehicle-treated well, representative single-agent-treated wells and a well containing the doublet are shown. All 4-well images belong to unique 12-well dishes. Representative images of three independent experiments. c In vivo dosage of imatinib, GDC-0994, and palbociclib at standard doses for animal use led to decreased AKT, P90RSK, and RB phosphorylation levels (targets of the kinases c-Kit, ERK, and CDK6, respectively) in MDA-MB-231-xenografted tumors after 24 h. The right panel shows total PNKP levels in wild-type (upper) and CRISPR PNKP MDA-MB-231 transfectant (lower panels) tumors. Scale bar, 50 μm
