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Fig. 4 | BMC Immunology

Fig. 4

From: Cyclosporine a directly affects human and mouse b cell migration in vitro by disrupting a hIF-1 αdependent, o2 sensing, molecular switch

Fig. 4

Cyclosporine A interferes with B cell hypoxia dependent CXCR4 hypo-responsiveness by destabilizing HIF-1 α. RAMOS cells were incubated at 19, 4 or 1% O2 levels, with some 1% samples being incubated with the indicated concentrations CyA. After 24 hours the cells were subjected to (a) chemotaxis assay or (b) lysates were prepared and westerns performed to measure HIF-1 α levels with actin being used as a loading control. Shown is a representative blot. (c) Quantitation of relative HIF-1 α levels from three separate experiments. Primary human (d) or mouse (e) B cells were incubated at 19, 4 or 1% O2 levels, with some 1% samples being incubated with the indicated concentrations CyA. After 24 hours a chemotaxis assay was performed. (f) RAMOS B cells, stably transfected with lentiviral vectors expressing either HIF-1 α shRNA (H), a scrambled non-specific control sequence shRNA (C) or parental non-transfected RAMOS cells (-), were incubated at 1% O2 levels with the indicated concentrations of CyA. After 24 hours the cells were subjected to the chemotaxis assay. ***P <0.001; **P <0.01; *P <0.05

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