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Fig. 3 | BMC Immunology

Fig. 3

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

Fig. 3

HIF-1 α is both necessary and sufficient to induce chemokine receptor hypo-responsiveness. 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 the indicated O2 levels for 24 hours and then 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. Additionally, RAMOS B cells, stably transfected with lentiviral vectors over-expressing HIF-1 α (H), expressing firefly luciferase (C), or parental non-transfected RAMOS cells (-), were incubated at the indicated O2 levels for 24 hours and then subjected to (d) chemotaxis assay or (e) 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. (f) Quantitation of relative HIF-1 α levels from three separate experiments. ***P <0.001; **P <0.01

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