Fig 1: Over-expression of POU3F2 antagonized miR-107 inhibitory effects on melanoma cells. a Graphic summary of cell migration rate in wound closure experiment. SH-4 cells were co-transfected with miRNAs and POU3F2 or GFP. An open wound was created in the centre of a confluent monolayer of transfected cells. The percentage of the remaining open wound was recorded every 24 h for 4 days. b Cell invasive potential of transfected cells was measured with CytoSelect™ 24-Well Cell Invasion assay with type I collagen based matrix. SH-4 cells were co-transfected with miRNAs and POU3F2 or GFP. The transfected cells were plated on the upper chamber of a tran-well system. 48 h later, the matrix was collected and the invaded cells were stained with CyQuant® GR dye and quantified in a fluorescence plate reader. c Colony formation study of SH-4 cells co-transfected with miRNAs and POU3F2 or GFP. The colony formation ability of the transfected cells was accessed by counting the number of colonies (more than 50 cells) in the culturing plates. The data were presented as mean ± SD of at least three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001 as compared with control)
Fig 2: Schematic diagram of proposed mechanism. miR-107 expression may regulate melanoma cell proliferation, migration, invasion and apoptosis by inhibiting the expression of POU3F2
Fig 3: POU3F2 is a downstream target of miR-107 in melanoma cells. a Schematic diagram of a conserved putative miR-107 targeting site in 3'UTR of POU3F2 (WT). The site mutations were introduced to miR-107 targeting site of 3'UTR (MUT). b Luciferase reporter assay study of the interaction between miR-107 and 3'UTR of POU3F2. SH-4 cells were transfected with luciferase reporter genes conjugated with either wildtype (WT) or mutant (MUT) 3'UTR of POU3F2. 24 h later, the cells were transfected further with either miR-107 mimic or scramble control. Transfected cells were cultured for 48 h and then harvested for the measurement of luciferase activities in a plate reader. c Western blotting analysis of POU3F2 in cells transfected with either miR-107 or control miRNA. The representative image from at least three independent experiments was shown. d The densitometric analysis of POU3F2 expression was shown. The data were presented as mean ± SD of at least three independent experiments: (*p < 0.05, **p < 0.01, ***p < 0.001 as compared with control)
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