Hypoxia-inducible factor 1 alpha limits dendritic cell stimulation of CD8 T cell immunity
Fig 4
HIF-1α expression in BMDC cultures promotes the production of immunosuppressive cytokines.
(a) Nitric oxide production by WT and HIF-1α-Tg BMDCs was indirectly quantified by measuring nitrite levels (Griess reaction) by ELISA. (b) Arginase I and HIF-2α protein levels in total WT and HIF-1α-Tg BMDC cell lysates. Cell lysates from the same samples were split across different gels and probed separately for arginase I and HIF-2α. β-tubulin was used as a protein loading control. (c) VEGF production by BMDCs was analyzed by ELISA. (d) Il10 expression in BMDCs was analyzed by qPCR before and after CpG-stimulation. (e) IL-10, (f) IL-6, (g) IL-12p70, (h) TNFα, and (i) TGFβ production by BMDCs was analyzed by ELISA. All data are representative of two to three independent experiments. Error bars represent SD of the mean. Statistical testing was performed using two-way ANOVA and Tukey’s test for multiple comparison; n.s.: not significant, n.d.: not detected (below limit of detection).