Skip to main content
Fig. 6 | Journal of Hematology & Oncology

Fig. 6

From: JAG1 overexpression contributes to Notch1 signaling and the migration of HTLV-1-transformed ATL cells

Fig. 6

Repression of JAG1 alters Notch signaling and wound healing in ATL lines. a RT-PCR was performed on JAG1 from cDNA derived from ATL cell line ATL55T infected with a lentivirus pSIH1-shRNA against JAG1. Cells infected with the empty vector expressing pSIH1-GFP were used as a control. The expression of JAG1 was normalized to GAPDH expression. RT-PCR was then performed on Hes-1, Hey-1, and VEGF from cDNA derived from ATL55T cells infected with pSIH1-shRNA against JAG1. Cells infected with the empty vector expressing pSIH1-GFP were used as a control. Real-time PCR was performed in duplicate, and samples were normalized to GAPDH expression. b Real-time PCR was performed on Hes-1 and Hey-1 on cDNA extracted from ATL55T and Jurkat cells incubated for 6 days with neutralizing antibody against JAG1 (3 μg/ml). Real-time PCR was performed in duplicate, and samples were normalized to GAPDH expression. c Cell proliferation was measured using XTT assay in ATL55T and Jurkat cells incubated for 6 days with neutralizing antibody against JAG1 (3 μg/ml). Results were plotted as mean ± standard deviation from at least two independent experiments. d PI staining was performed to study cell death in ATL55T and ATL43T cells incubated for 6 days with neutralizing antibody against JAG1 (3 μg/ml). Media with antibody was replaced every 3 days, and fresh antibody was added (3 μg/ml). Cells were analyzed for apoptosis by FACS analysis. Bar diagrams representing the FACS results are provided. e Wound healing was performed on confluent ATLT cells treated with 3 μg/ml of JAG1-neutralizing antibody for 72 h. After 72 h, the p1000 tip was used to scratch the plate. The plate was then washed, and fresh media with neutralizing antibody was added. After 48 h, the cells were fixed and images were taken with a × 40 objective lens. Bar diagrams representing the wound healing results are provided

Back to article page