Fig. 6

JAK2 inhibitors reverse the expression of the PITPNC1-regulated transcriptome and synergize with Sotorasib. A Connectivity Map (CMap) analysis of dPITPNC1 GS obtained in A549 cells. Perturbagen classes with mean connectivity scores > 90% are displayed. Each dot represents and individual drug included in the specific class. B Fedratinib (Fedra) IC50 index in a panel of LUAD and PDAC cell lines treated with the drug for 5 days. C Western blots of MYC and LC3-I/LC3-II in H358, H2009, HPAFII and MiaPaca2 treated with DMSO (C), and 2 or 10 μM of Fedra 48 h. Twenty μg of protein were loaded per sample and HSP90 was used as loading control. D Heatmaps of H1792, H2030, H358, H23 and MiaPaca2 cell viability percentage after treatment for 5 days with different concentrations of Sotorasib (Soto) and Fedra, individually or in combination. E and F Effects of Soto and Fedra combination on cell viability of mut KRAS LUAD cells (H1792 and H358) grown in 3D culture conditions, 5 days after drug treatment. Soto: 60 nM; Fedra: 1 μM. (Dunnett’s multiple comparison test). G Western blots of KRAS, pERK1/2, ERK1/2, pSTAT3, STAT3, HSP90, caspase 3, cleaved caspase 3 and GAPDH in H358 and MiaPaca2 cell lines treated with vehicle (Ctrl), 20 nM Soto, 1 μM Fedra, or both (Combo) for 48 h. Twenty μg of protein were loaded per sample. HSP90 and GAPDH were used as loading controls. H Representative image and quantification of clonogenic capacity of H2030 (Soto: 5 nM; Fedra: 0.5 µM), H358 (Soto: 5 nM; Fedra: 0.25 µM), H23 (Soto: 5 nM; Fedra: 0.5 µM) H1792 (Soto: 20 nM; Fedra: 0.5 µM) and MiaPaca2 (Soto: 20 nM; Fedra: 0.5 µM) cells treated with the indicated drugs and concentrations for 10 days, (Dunnett’s multiple comparison test). I Heatmaps showing cell viability percentage of Soto-resistant (SR) H23 and H358 cell lines treated with different concentrations of Soto and Fedra, individually or in combination. J Synergistic score (Bliss score) heatmaps of Soto-resistant (SR) H23 and H358 cells treated for 5 days as indicated