Fig 2: Extended cMet/PI3K pathway model. (A) Schematic representation of the model structure including mCherry-AKT. (B) Dynamics of the phosphorylation of cMet, endogenous AKT, and mCherry-AKT in the Hepa1_6-D8 and E2 clones stimulated with 40 ng/ml HGF. The means of the indicated number of biological replicates are represented as dots with the standard deviation as error bars. The model trajectories are depicted as lines and the corresponding Chi-square values are given.

Fig 3: PI3K pathway activation in ECs leads to enhanced cell cycle progression AImmunoblot showing the activation of PI3K/AKT pathway (by assessing the levels of p-AKT Ser473) of ECs 24 h after induction of Pik3caH1047R expression.BVolcano plot of genes analysed by RNAseq in ECs upon Pik3caH1047R expression. Differentially expressed genes (DEGs) (P adj < 0.05) in Pik3caH1047R over Pik3caWT ECs at 24 h (grey, unchanged; orange, downregulated; purple, upregulated). n = 4 biological replicates per genotype.CPathway enrichment analysis showing most represented biological functions altered upon Pik3caH1047R expression in ECs.DImmunoblot showing the activation of PI3K/AKT pathway of Pik3caWT and Pik3caH1047R ECs after 2 h starvation (-) and 30 min stimulation with 20% foetal bovine serum (FBS) (+).E, F(E) Cell cycle analysis by flow cytometry of Pik3caWT and Pik3caH1047R ECs after cultured either without serum (-) or 20% serum (+) for 24 h. (F) Percentage of proliferative ECs (in S and G2 M phases) from Fig 1E. Error bars are SEM. n = 3 biological replicates per genotype. Statistical analysis was performed by nonparametric Mann–Whitney test. **P < 0.01 and ****P < 0.0001 were considered statistically significant. Source data are available online for this figure.

Fig 4: Energy/glucose homeostasis and insulin action in SMAKO mice. Body weight (A), fat mass (B), and lean mass (C) (?, 3–28 weeks; n = 19/group). D: Muscle weights. With the exception of heart, data represent the average weight for both muscles from each mouse (?, 28 weeks, fasted 12 h; n = 19/group). EDL, extensor digitorum longus; Gas, gastrocnemius; Hrt, heart; Plant, plantaris; Quad, quadriceps; Sol, soleus; TA, tibialis anterior. E: RER using a Comprehensive Lab Animal Monitoring System (CLAMS) (?, 11 weeks, weight-matched; n = 4/group). F: GTT at 19 weeks with 1.875 g/kg glucose i.p. (?, fasted 12 h; n = 17–20/group). G: ITT at 20 weeks with 1.1 units/kg insulin i.p. (?, fasted 4 h; n = 17–20/group). H–K: Insulin signaling studies: mice were fasted for 12 h, injected i.p. with saline or insulin at 10 units/kg body weight, and killed 10 min thereafter (?, 28 weeks, tibialis anterior; n = 5–7/group). Representative immunoblots (H) and associated quantification of stoichiometric phosphorylation of Akt pS473/Akt total (I), Akt pT308/Akt total (J), and total Akt/Ran GTPase (RAN) control (K). Fold change in response to insulin treatment is indicated above the black bars. For overall effects having P < 0.05: D, diet; G, genotype; N, nutritional status (i.e., fasting/refeeding); T, treatment (i.e., with insulin). For clarity, only overall effects are shown.

Fig 5: Proposed model of Integrin Tie2 cooperative signaling in endothelial cell regulation.(A) Based on previous studies, it is known that Ang-1 clusters Tie2 at cell-cell junctions in endothelial cells while providing pro-survival signals through the AKT and Survivin signal transduction pathway [30,46,47]. Alternatively, Tie1 may associate with integrins, including either aVß3 or a5ß1, which are preferentially localized at the cell interface with the basement membrane containing extracellular matrix components. A link between Tie1 and the extracellular matrix could explain a role for Tie1 in vessel integrity and sheer stress that has previously been described [43–45]. (B) When Tie2 is not at junctions, it is likely capable of associating with integrins. Activation of integrin-associated Tie2 via Ang-1 and fibronectin may enrich signaling through MAPK to further facilitate endothelial cell migration and proliferation.