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Fig. 6 | BMC Cancer

Fig. 6

From: TNF-alpha promotes lymphangiogenesis and lymphatic metastasis of gallbladder cancer through the ERK1/2/AP-1/VEGF-D pathway

Fig. 6

The TNF-α - VEGF-D axis promoted the tube formation of human dermal lymphatic endothelial cells (HDLECs) in vitro. a, b Construction of a NOZ cell line and a GBC-SD cell line stably expressing lentiviral VEGF-D shRNA and a green fluorescent protein sequence. The cells were observed under a fluorescence microscope with bright or blue light. c, d VEGF-D mRNA and protein expression of NOZ or GBC-SD cells stably transfected with LV-siVEGF-D were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. GAPDH served as an internal control. e, f, g, h DiI-labeled HDLECs (emit red fluorescence) were cocultured with the three NOZ (or GBC-SD) cell lines and were treated with TNF-α (50 ng⁄ mL) for 5 h. HDLEC tube formation was observed under fluorescence microscopy, and the tube number was counted. (*P < 0.05; **P < 0.01; ***P < 0.001)

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