Fig. 5

PITPNC1 controls mTOR localization to prevent autophagy. A Gene Ontology analysis of the upregulated PITPNC1 gene set (uPITPNC1 GS). B and C SESN1, SESN2 and SESN3 expression levels in A549 (B) and HPAFII (C) cell lines were measured by qPCR. Cells were virally infected to express a control (GFPsh) or a PITPCN1 shRNA (sh6 and sh7) (Dunnet’s multiple comparison test). GAPDH was used as housekeeping gene. D and E mTOR/LAMP1 colocalization analysis by immunofluorescence in A549 (D) and HPAFII (E) PITPNC1-depleted cells. F and G Quantification of mTOR/LAMP1 Mander’s overlap coefficient (MOC) in A549 (F) and (G) of D and E (Dunnett’s multiple comparison test). H Lysosomes per cell and average lysosomes size in A549 of D (Dunn’s multiple comparison test). I Lysosomes per cell and average lysosomes size in HPAFII of E (Dunn’s multiple comparison test). J Western blots of LC3-I and LC3-II protein levels in a LUAD (n = 3) and PDAC (n = 3) cell lines expressing a shRNA control (C) or two PITPNC1 shRNAs (sh6 and sh7). Twenty μg of protein were loaded per sample and HSP90 was used as loading control. K Western blots of protein levels of LC3-I and LC3-II in a LUAD (n = 3) and PDAC (n = 1) cell lines expressing a shRNA control (C) or two MYC shRNAs (sh42 snd sh89). Twenty μg of protein were loaded per sample and HSP90 was used as loading control. L Proposed model for the role of PITPNC1 in KRAS-driven LUAD and PDAC