Fig 2: Western blots of the proteins of PDGF-B signaling pathwayThe different expressions from three individual patients are presented for the selected proteins. PDGF-B, PDGFR-ß, eIF4E and p70s6K were increased in the gastric cancer tissues, compared to normal gastric tissues, Akt expression is increased in two groups, ERK1/2 expression was not significantly altered. The phosphorylation and activation status of Akt, ERK, p70S6K and eIF4E were observed significantly increased. All the experiments were repeated at least three times, N represents normal tissue and T represents tumor tissue.

Fig 3: Deletion of GPR44 worsens AML by activation of the PI3K/AKT/mTOR pathway(A–D) Expression of genes including PI3K (Pik3ca, Pik3cb, Pik3cd) (A), AKT (Akt1, Akt2) (B), PKC (Prkca, Prkcb, Prkcg) (C), and PKA (Prkacg) (D) assessed by qPCR analysis in WT or Gpr44-/- AML cells isolated from spleens of 2 AML-recipient mice. Data were normalized to WT AML cells and Gapdh expression (n = 8–10 biological replicates in each group).(E) Western blot showing the expression of Phospho-PI3K, PI3K, Phospho-mTOR, mTOR, Phospho-AKT, AKT, Phospho-P70-S6K, P70-S6K, and ß-actin in purified 1° WT and Gpr44-/- AML cells.(F) Western blot showing the expression of Phospho-PTEN, PTEN, Phospho-4EBP1, 4EBP1, and ß-actin in WT or Gpr44-/- AML cells isolated from spleens of 2° AML-recipient mice (n = 5 biological replicates in each group).(G) Viability of purified 1 Gpr44-/- AML cells treated with 150 µM ARN509, 10 µM LY294002, 10 nM Wortmannin, 10 nM Sapanisertib, 250 nM Torin1 for 24 h. Data were normalized and compared with vehicle treatment.(H and I) Flow cytometric analysis of Phospho-P70-S6K (H) and Phospho-4E-BP1 (I) in CD45.1+ 1° Gpr44-/- AML cells treated with 10 µM LY294002, 10 nM Wortmannin, 10 nM Sapanisertib, 250 nM Torin1 for 24 h. MFI was summarized (n = 4).(J) Counts of CFUs of purified 1° Gpr44-/- AML cells treated with 10 µM LY294002, 10 nM Sapanisertib, and 2.5 µM LY2584702 in methylcellulose medium (2,500 cells/well, n = 3 for technical replicate) CFUs were counted on day 8.(K) Representative image of colony growth in (J). Scale bar, 100 µm.Data shown are mean ± SEM per group; *p < 0.05, ** p < 0.01.