Fig. 4

PITPNC1 loss induces a G1 phase arrest linked to MYC downregulation. A Gene Ontology analysis of the downregulated PITPNC1 gene set (dPITPNC1 GS). B and C Cell cycle analysis by EdU labelling in the human LUAD A549 and H2009 (B), and PDAC HPAFII and Panc1 (C) cell lines after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh). (Bonferroni´s multiple comparison test). D MYC mRNA expression in A549, H2009 and H1792 LUAD and PDAC PATU8902, Panc1 and HPAFII cell lines expressing a specific shRNA (sh6 or sh7) compared to control (GFPsh) (Dunnet´s multiple comparison test). E MYC protein expression in the A549, H2009 and H1792 LUAD cell lines after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh). Twenty μg of protein were loaded per sample. β-TUBULIN was used as loading marker. F MYC protein expression in Panc1, HPAFII and MiaPaca2 PDAC cell lines after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh). Twenty μg of protein were loaded per sample. β-TUBULIN was used as loading marker. G E2F1 and p27 protein expression in the A549, H2009 and H1792 LUAD cell lines after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh). Twenty μg of protein were loaded per sample. β-TUBULIN was used as loading marker. H E2F1 and p27 protein expression in the PATU8902, HPAFII and MiaPaca2 PDAC cell lines after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh). Twenty μg of protein were loaded per sample. HSP90 was used as loading marker. I MYC and PITPNC1 protein expression in H2009 and PATU8902 MYC-overexpressing cells after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh) and treated with DMSO or MG132 (10 μM, 6 h). Twenty μg of protein were loaded per sample. HSP90 was used as loading marker. J AURKA and PLK1 protein expression in A549, H2009 and H1792 LUAD cell lines after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh). Twenty μg of protein were loaded per sample. HSP90 was used as loading marker. K AURKA and PLK1 protein expression in Panc1, HPAFII and MiaPaca2 PDAC cell lines after PITPNC1 knockdown with a specific shRNA (sh6 or sh7) compared to control (GFPsh). Twenty μg of protein were loaded per sample. HSP90 was used as loading marker. L MYC protein levels in A549 and H1792 LUAD and Panc1 and HPAFII PDAC cell lines treated with DMSO, PLK1i (BI2536, 50–100 nM, 48 h), or both PLK1i plus proteasome inhibitor (MG132, 10 μM 6 h). Twenty μg of protein were loaded per sample. HSP90 was used as loading marker