Fig. 1
PITPNC1 is upregulated in KRAS-mutated LUAD and PDAC and predicts poor survival. A Heatmap of upregulated genes in The Cancer Genome Atlas (TCGA) LUAD data set comparing expression profiles of wt and mut KRAS LUAD patients. B PITPNC1 mRNA expression levels in normal lung (N), wild type (wt) and mutant (mut) KRAS LUAD. Mut vs wt KRAS (p < 0.0001) or vs N (p < 0, 0001). C PITPNC1 gene amplification percentage (GISTIC2 analysis) in mut and wt KRAS LUAD samples, or both (p = 0.815). D Kaplan–Meier survival analysis of LUAD patients, stratified based on KRAS status and PITPNC1 expression. Data from TCGA database: wt KRAS (Log-rank test p = 0.96) and mut KRAS (Log-rank test p = 0.04). E Kaplan–Meier survival analysis of PDAC patients stratified by PITPNC1 expression. Data from ICGC database (Log-rank test p = 0.027). F Western blot of PITPNC1 and KRAS expression in H2126 and H6C7 cells, expressing a control (LacZ) or overexpressing KRAS (wt KRAS4B or mut KRASG12D, G12C or G12V). Twenty μg of protein were loaded per sample. HSP90 and β-TUBULIN were used as loading markers. G Western blot of PITPNC1 and KRAS expression in A549, H2009, PATU8902 and HPAFII cells, expressing a control (GFPsh) or an inducible KRAS shRNA (KRASsh) (activated by 1 µg/ml doxycycline). Twenty μg of protein were loaded per sample. HSP90 were used as loading markers. H Western blot of PITPNC1 expression in A549, H2009 and HPAFII cells treated for 24 h with pharmacologic inhibitors: trametinib (MEKi, 0.5 μmol/L), BIX02189 (MEK5i, 10 μmol/L), SP600125 (JNKi, 10 μmol/L) or GSK2126458 (PI3Ki, 0.1 μmol/L). Twenty μg of protein were loaded per sample. β-TUBULIN was used as loading marker. I Western blot of PITPNC1 and KRAS expression in Kraslox/lox MEFs transduced with different human HA-tagged KRAS mutants (G12C, G12D, G12V, G12R, G12S, G13D and Q61H). 4OHT: 600 nM. J PITPNC1 mRNA expression levels in no loss of heterozygosity (no LOH) and loss of heterozygosity (LOH) TCGA LUAD patients. no LOH vs LOH (p = 0.047)