Fig. 6

MSI2a binds to TP53INP1 mRNA and increases TP53INP1 mRNA stability and protein expression. a Schematic map of construction of the 3′-UTR reporter constructs for TP53INP1 that were used for a dual-luciferase assay. TP53INP1 mRNA stability curves plotted using qPCR expression versus time. b-d Luciferase reporter assay. (b) MDA-MB-231 cells transfected with MSI2a plasmids and (c) Hs-578 T cells transfected with MSI2a siRNA were treated with actinomycin D (5 mg/mL) for the indicated periods of time. (d) MDA-MB-231 cells were cotransfected with MSI2-overexpressing plasmids, Renilla luciferase 3′-UTR constructs, and a control firefly luciferase control vector. e Hs-578 T cells were cotransfected with MSI2a-specific siRNAs, Renilla luciferase 3′-UTR constructs, and a control firefly luciferase control vector. f Luciferase reporter assay. MDA-MB-231 and Hs-578 T cells were cotransfected with MSI2a-overexpressing plasmids, Renilla luciferase, 3′-UTR constructs, and a control firefly luciferase control vector. Renilla luciferase activity following MSI2a overexpression was determined. g Luciferase reporter assay. Schematic map of construction of the MSI2a-Mut plasmids. MDA-MB-231 cells were cotransfected with MSI2a or MSI2a-Mut plasmids, Renilla luciferase, 3′-UTR constructs, and a control firefly luciferase control vector. Renilla luciferase activity was determined. The data were normalized to firefly luciferase activity. *p < 0.05, **p < 0.01, ***p < 0.001