Fig 2: Effect of anti-DLL4 and DAPT on the expression of apoptosis-related genes and the body weight of mice in orthotopic gastric tumor models.(A) Representative immunohistochemistry staining for BAX, Bcl-2, and P53 in the tumors of each group. 400× magnification. (B) AOD of immunohistochemistry staining of tumor samples in each group. *P < .05, when compared to the MB+UTMD control. (C) No significant anti-DLL4– and DAPT-related body weight loss was observed in the treated mice.

Fig 3: Effect of anti-DLL4 and DAPT on apoptosis and cell cycle in gastric cancer cells.(A to D) Cells were stained with Annexin V and PI, following which cell apoptosis in each group was detected by flow cytometry. (E to H) Cell cycle phase distribution was measured by staining DNA with PI followed by flow cytometry. (I and J) Cell apoptosis rate and cell cycle distribution in each group were analyzed. *P < .01, when compared with control group.

Fig 4: Effect of anti-DLL4 and DAPT on tumor growth in orthotopic gastric tumor models.Tumor growth was monitored and quantified by real-time whole-body fluorescence imaging. Final tumor weight was determined at autopsy. (A) In vivo whole-body fluorescence imaging of tumor before treatment and 18 days after treatment in each group. (B) Markedly increased necrogenous reaction in the parenchymal of tumor in the combined treatment group was observed by H&E staining. (C) Tumor growth curves in each group. (D) Final tumor weights at the end of the study period in each group. *P < .01, when compared to MB+UTMD control. #P < .01, when compared to MB+DAPT+UTMD group.

Fig 5: DLL4 protein expression in SGC-7901-GFP cells.Membranous expression pattern of DLL4 protein in SGC-7901 cells detected by the rosette formation test with anti-DLL4 antibody conjugated microbubbles. (A) No rosette formation observed with SGC-7901 cells treated with unconjugated microbubbles. (B) Rosette-like structure formed with SGC-7901 cells treated with anti-DLL4 antibody conjugated microbubbles. (C) Rosette formation blocked by adding anti-DLL4 antibody prior to SGC-7901 cells being treated with anti-DLL4 antibody conjugated microbubbles. Bars = 50 µm. DLL4 expression in SGC-7901 cells was detected by immunofluorescence. (D) Red fluorescence showing DLL4 expression in SGC-7901 cells. (E) DAPI showing SGC-7901 nuclei. (F) Merged image of A and B.