Figure 2.

Cell-Autonomous KRAS^G12D Phosphoproteome

(A) KRAS^WT and KRAS^G12D PDA cell lysates were isobarically labeled with tandem-mass tags (TMT) (126–131 mass-to-charge ratio [m/z]), mixed, and subjected to automatic phosphopeptide enrichment (APE) (n = 5). TMT-phosphopeptides were analyzed by high-resolution LC-MS/MS and normalized to total protein level changes.

(B) KRAS^WT and KRAS^G12D phosphoproteomes cluster in PCA space.

(C) Statistical regulation of the PDA KRAS^G12D cell-autonomous phosphoproteome (n = 5, two-tailed t test, Gaussian regression). Cell-autonomous enriched phospho-motifs shown.

(D) PDA cell-autonomous regulation of 18 intracellular signaling nodes following KRAS^G12D induction across 48 hr (n = 3) in PCA space.

(E) KRAS^WT and KRAS^G12D PDA cells treated ±MEK and AKT inhibitors analyzed by multivariate phosphoproteomics. KRAS^G12D cell-autonomous PDA phosphoproteomic state requires active MEK and is independent of AKT activity.

See also Figures S2, S3, and Data S1.