Fig 2: GRB2 was regulated by BMPR2 in PDAC. (A) Heat map of protein spectrum analysis. (B) Volcano diagram analysis of protein spectrum analysis. (C) PPI network analysis after down-regulation of BMPR2 in MiaPazCa-2 cells. (D) Expression of GRB2 mRNA in PDAC tissues (n=35). (E) Representative western blot analysis of expression of GRB2 in normal pancreas and PDAC tissues. (F) Analysis of expression of GRB2 in normal pancreas and PDAC tissues (n=35). (G) Overall survival rates of pancreatic cancer patients with low or high expression of GRB2. (H) Correlation of BMPR2 and GRB2 in PDAC tissues (data from TCGA database). (I,J) Expression and quantitative analysis of GRB2 after inhibition of BMPR2 in MiaPaCa-2 and PANC-1 cells. ***, P<0.001. PPI, Protein-protein interaction; BMPR2, bone morphogenic protein receptor 2; PDAC, pancreatic ductal adenocarcinoma

Fig 3: LDN193189 inhibited PDAC growth in vivo. (A) Representative bioluminescent images of orthotopic pancreatic cancer models during the treatment. (B) Bioluminescent densities during the treatment. (C) Tumor images at the end of the treatment, n=5. (D) Representative IHC images showing the expression of BMPR2, GRB2 and Ki67 of tumors. (E) Western blot analysis of the orthotopic tumors. (F) Tumor growth curves of PDX during the treatment. (G) Image of tumors excised from PDX mouse models at the end of the treatment, n=5. (H) Tumor weight analysis of PDX models. (I) Western blot analysis of the PDX tumors. (J,K) Tumor growth after knockdown of BMPR2 by shRNA (n=5). Scale bar represents 1 cm. The data are shown as the mean ± SD. ***, P<0.001. IHC, immunohistochemistry; BMPR2, bone morphogenic protein receptor 2; PDX, patient-derived xenograft; PDAC, pancreatic ductal adenocarcinoma.

Fig 4: Down-regulation of BMRP2 induced suppression of PI3K/AKT signaling pathway. (A) Bubble chart of protein spectrum analysis. (B) Enrichment plot analysis of PI3K/AKT signaling pathway. (C) Level of p-PI3K and p-AKT in pancreatic cancer cells after down-regulation of BMPR2. (D) Level of p-PI3K and p-AKT in pancreatic cancer cells after down-regulation of GRB2.

Fig 5: Inhibition of BMPR2 suppressed proliferation of pancreatic cancer cells. (A) Representative images of IHC results. (B) Analysis of BMPR2 expression in normal pancreas and PDAC tissues. (C) Correlation of BMPR2 expression level and overall survival of PDAC patients. (D) Expression level and quantitative analysis of BMPR2 in PDAC cell lines and a normal ductal epithelial cell line, HPNE. (E) Verification of two sequences of shRNA in down-regulating BMPR2 in two PDAC cell lines. (F) Proliferation of MiaPaCa-2 and PANC-1 cells after down-regulation of BMPR2. (G,H) Colony formation assay and quantitative analysis of MiaPaCa-2 and PANC-1 cells after down-regulation of BMPR2. Scale bar: 1 cm. (I) Cell cycle distribution of MiaPaCa-2 and PANC-1 cells after down-regulation of BMPR2. (J) G2/M related proteins detection after knockdown of BMPR2. IHC, immunohistochemistry; BMPR2, bone morphogenic protein receptor 2; PDAC, pancreatic ductal adenocarcinoma. *, P<0.05; **, P<0.01; ***, P<0.001.