Fig. 7

mt-e-CAFs impair cancer cell response to standard-of-care gemcitabine/Abraxane in vitro. a Representative images and quantification of Ki67 staining and of b cleaved-caspase 3 (CC3) staining of fl-CCs and mt-CCs invading into fl-e-CAF matrices upon treatment with control or with gemcitabine (100 nM)/Abraxane (100 nM) for 72 h. Scale bar: 50 μm, n = 3 biological repeats with three technical replicates per biological repeat. c Representative images and quantification of Ki67 staining and of d CC3 staining of fl-CCs and mt-CCs invading into mt-e-CAF matrices upon treatment with control or with gemcitabine (100 nM)/Abraxane (100 nM) for 72 h. Scale bar: 50 μm, n = 3 biological repeats with three technical replicates per biological repeat. e Schematic representation of the CDK1-FRET biosensor (adapted from 32) and representative mCerulean lifetime maps showing high CDK1 activity or low CDK1 activity as assessed via FLIM-FRET imaging of the CDK1-FRET biosensor. f Schematic representation of co-seeding of CAFs and cancer cells expressing the CDK1 biosensor on dishes coated with native fibrillar collagen, treatment with gemcitabine/Abraxane and FLIM-FRET imaging of CDK1 activity in cancer cells. g Representative mCerulean lifetime maps and h quantification of mCerulean lifetimes and i. CDK1 activity in fl-CCs cultured alone, cultured with fl-e-CAFs or with mt-e-CAFs and treated with control or with gemcitabine (100 nM)/Abraxane (100 nM) for 24 h. Scale bar: 100 μm, n = 3 biological repeats. j Representative mCerulean lifetime maps and k quantification of mCerulean lifetimes and l CDK1 activity in mt-CCs cultured alone, cultured with fl-e-CAFs or with mt-e-CAFs and treated with control or with gemcitabine (100 nM)/Abraxane (100 nM) for 24 h. Scale bar: 100 μm, n = 3 biological repeats, >100 cells analyzed per condition. Individual data points are presented with mean values and SEM. *p < 0.05