Figure 6
Cell viability in response to In1-ghrelin overexpression and silencing in tumoral pituitaries.
Effect of 24-, 48- and/or 72-h overexpression of ghrelin (Ghr) and/or In1-ghrelin (In1) on cell viability in human GH-omas (A; n = 4–11), NFPAs (B; n = 2–3) and ACTH-omas (C; n = 3) primary cell cultures, determined by alamar-blue reduction. Overexpression was induced by transfection with specific expression vectors containing the appropriate CDS of each variant. Effect of 24- and 48-h silencing of In1-ghrelin (In1) expression on cell viability of individual human GH-omas (D; n = 2) and NFPAs (E; n = 2) primary cultures, determined by alamar-blue reduction. In1-ghrelin silencing was induced by transfection with specific siRNAs. Representative validation by qPCR of In1-ghrelin (black bar) and ghrelin (grey bar) overexpression (F) and silencing (G) of In1-ghrelin, demonstrating a decrease of In1-ghrelin mRNA expression (black bar) but no significant decrease of ghrelin mRNA expression (grey bar). Data are expressed as percent of control vectors (A, B, C. Mock; set at 100%) and control random siRNA (D, E. Scr: Scramble; set at 100%) within experiment, respectively. Values represent the mean ± S.E.M. Asterisks (* p < 0.05, ** p < 0.01; *** p < 0.001) indicate data that significantly differ from mock or scramble controls.
