Journal: Journal of Cellular and Molecular Medicine

Article Title: Inhibition of miR‐148a‐3p resists hepatocellular carcinoma progress of hepatitis C virus infection through suppressing c‐Jun and MAPK pathway

doi: 10.1111/jcmm.14045

Figure Lengend Snippet: miRNA mimics and inhibitor, c‐Jun siRNA sequence

Article Snippet: SiRNA 1 and siRNA 2 for silencing c‐Jun (si‐ c‐Jun ‐1 and si‐ c‐Jun ‐2) were purchased from Ribobio.

Techniques: Sequencing

Journal: BMC Cancer

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

doi: 10.1186/s12885-016-2259-4

Figure Lengend Snippet: TNF-α upregulated VEGF-D expression and VEGF-D promoter activity downstream of the ERK1/2/AP-1 pathway. a , c The effect of the TNF-α⁄AP-1 signaling pathway on the promoter activity and protein expression of the VEGF-D gene. Transfection with AP-1 siRNA effectively knocked down the expression of AP-1 and p-AP-1 in both NOZ and GBC-SD cells. The protein level and promoter activity of VEGF-D were accordingly reduced irrespective of treatment with TNF-α. b , d The effect of inhibition of MAPK pathway members on the protein expression and promoter activity of VEGF-D. When treated with SP600125 (10 μM), SB203580 (20 μM) or PD98059 (50 μM), the expression of AP-1 and p-AP-1 in both NOZ and GBC-SD cells were reduced. However, the protein expression and promoter activity of VEGF-D were significantly reduced only in the PD98059-treated group. * P < 0.05

Article Snippet: The AP-1 (c-Jun) siRNA interference sequence has been described previously [ ] (named siAP-1, sense: 5′-GAUGGAAACGACCUUCUAUdTdT-3′, anti-sense: 5′-AUAGAAGGUCGUUUCCAUCdTdT-3′), and the non-targeting control (named siNC) were synthesized chemically by GenePharma Co., Ltd. (Suzhou, China).

Techniques: Expressing, Activity Assay, Transfection, Inhibition

Journal: BMC Cancer

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

doi: 10.1186/s12885-016-2259-4

Figure Lengend Snippet: 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)

Article Snippet: The AP-1 (c-Jun) siRNA interference sequence has been described previously [ ] (named siAP-1, sense: 5′-GAUGGAAACGACCUUCUAUdTdT-3′, anti-sense: 5′-AUAGAAGGUCGUUUCCAUCdTdT-3′), and the non-targeting control (named siNC) were synthesized chemically by GenePharma Co., Ltd. (Suzhou, China).

Techniques: In Vitro, Stable Transfection, Expressing, shRNA, Sequencing, Fluorescence, Microscopy, Transfection, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control, Labeling