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. 2020 Apr 18;2(1):vdaa047. doi: 10.1093/noajnl/vdaa047

Figure 3.

Figure 3.

MiR-93-induced changes in GBM-derived cytokine secretion are elicited via direct targeting of LIF, IL-6, G-CSF, Map3K2, and HIF-1α. (A) Luciferase reporter activity of cells co-transfected with constructs containing the 3′ UTR of COX2 (n = 3), CXCL5 (n = 3), LIF (n = 4), G-CSF (n = 5), IL-6 (n = 5), HIF-1α (n = 9), or Map3K2 (n = 11) and either miR-93 or scrambled control. All Luciferase experiments were performed in triplicates; ns = not significant, **P < .01. (B) Analysis of GBM cells incubated in 5% O2 and stimulated with IL-1 after siRNA-mediated double knockdown of HIF-1α and Map3K2 (DKO). Left panel: qRT-PCR analysis of COX2 and CXCL5 (n = 8); **P < .01. Right panel: Knockdown efficiency of HIF-1α and Map3K2 siRNA transfection (qRT-PCR, n = 3, **P < .01). (C) Left panel: CXCL5 protein expression after double knockdown of HIF-1α and Map3K2 as measured by ELISA; n = 6, *P < .05. Middle and right panel: COX-2 protein expression after double knockdown of HIF-1α and Map3K2, measured by immunofluorescence and SDS-PAGE. A typical example of 3 experiments is shown.