Figure 1
From: STAT3 governs the HIF-1α response in IL-15 primed human NK cells
IL-15 priming induces HIF-1α production only partially through mTORC1. (A–D) NK cells were incubated under normoxia (N, 21% O2) or hypoxia (H, 1% O2) for 16 h, primed with IL-15 for additional 6 h in the presence or absence of rapamycin (rapa) and the proteasome inhibitor MG132 as indicated. (A) Bead array analysis of pRPS6Ser235/236. Median fluorescence intensities (MFI) are represented as mean values ± SD (n = 4). Statistical significance was not reached when applying the Wilcoxon signed-rank test. (B) Immunoblotting with antibodies against HIF-1α and β-actin (loading control). The blot reflects one representative experiment of three performed. (C) RT-PCR analysis. Linearized quantities (2−ΔCt) are represented as mean values ± SD (n = 6). *p < 0.05, Wilcoxon signed-rank test. (D) Anti-HIF-1α and anti-β-actin immunoblot analysis. The blot reflects one representative experiment of two performed. (E) NK cells were incubated for 4 h with DMOG (chemical hypoxia) and with or without IL-15. Where indicated, cytokine-treated cells were cultured for additional time periods with cycloheximide (CHX) or vehicle (DMSO). Cultures were subjected to anti-HIF-1α and anti-β-actin immunoblot analysis. The blot reflects one representative experiment of three performed.
