Figures & data
Table 1 Clinical data of 70 patients with osteosarcoma
Table 2 Primer sequences
Figure 1 (A) Tumor cells were negative for Notch3 in case 12 (left); tumor cells were positive for Notch3 in case 44 (right) (EnVision ×400, scale 50 µm); (B) Kaplan–Meier survival curves and log-rank test for cumulative survival of the Notch3+ group and Notch3− group; P=0.017.
Table 3 Cox regression analysis
Figure 2 Changes in the Notch3 mRNA (A) and protein (B) expressions after lentiviral transfection.
Note: *P<0.05; **P<0.01.
Figure 3 (A) Effect of Notch3 knockdown on the proliferation of osteosarcoma cells in vitro. (B) Effect of Notch3 knockdown on the cell cycle of osteosarcoma cells. (C) Results of wound healing assay after Notch3 knockdown in osteosarcoma cells. Notch3 knockdown attenuated osteosarcoma cell migration. (D) Transwell invasion assay after Notch3 knockdown in osteosarcoma cells. Notch3 knockdown attenuated osteosarcoma cell invasion.
Note: *P<0.05; **P<0.01; ***P<0.001.
Figure 4 Notch3 knockdown inhibited Hes1 and MMP7 mRNA (A) and protein (B) expressions.
Note: *P<0.05; **P<0.01.
Abbreviation: MMP7, matrix metalloproteinase 7.
Figure 5 (A, B) Changes in the Hes1 mRNA and protein expressions after siRNA in osteosarcoma cells. (C, D) Hes1 knockdown reduced MMP7 expression.
Note: *P<0.05; **P<0.01.
Abbreviation: MMP7, matrix metalloproteinase 7.
Figure 6 (A) Gross pictures of pulmonary metastases in nude mice. (B) Histological pictures of pulmonary metastases in nude mice. (C) Effect of decreased Notch3 on the number of pulmonary metastases. (D) Differences in Notch3, Hes1, and MMP7 RNA expression between the control group and the Notch3 knockdown group in nude mice.
Note: *P<0.05; **P<0.01; ***P<0.001.
Abbreviation: MMP7, matrix metalloproteinase 7.
Table 4 Pulmonary metastasis rates in nude mice transplanted with MTT and U2OS cells
Figure S1 Jadded1+ tumor cells in case 48.
Figure S2 Jadded2− tumor cells in case 8.
Figure S3 DLL4+ tumor cells in case 36.
Table S1 Expression of 4 antigens in 70 osteosarcoma cases