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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: Overexpression of hypoxia-inducible factor 1 alpha improves immunomodulation by dental mesenchymal stem cells

Fig. 5

HIF-1 overexpression in MSCs confers resistance to NK cell-mediated lysis. Natural killer (NK) cells were cultured in media alone (black bars) or with rhIL-12 and rhIL-15 in the absence (white bars) or presence of green fluorescent protein (GFP)-mesenchymal stem cells (MSCs; light grey) and hypoxia-inducible factor (HIF)-MSCs (dark grey). After 36 h of culture, expression of CD107a (a), intracellular interferon (IFN)-gamma (c) and the indicated molecules (d) was analyzed in NK cells by flow cytometry. MSCs were excluded from the analysis by CD90 expression. b MSC-NK cell co-cultures were performed at the indicated MSC:NK cell ratios in presence of rhIL-12 and rhIL-15. After 36 h late apoptosis of MSCs was determined by CD90 expression and propidium iodide (PI) incorporation. A representative experiment (b and c) and mean ± SD of three independent experiments (a and d) are shown. One-way ANOVA, *p ≤ 0.05, GFP-MSCs and HIF-MSCs versus no MSCs. e, f MSCs were cultured until 80% confluence was reached, and then culture medium was replaced by fresh medium alone (e) or supplemented with IFN-gamma (f). After 15 h, cells were harvested and expression of the indicated genes was determined by qPCR. GNB2L1 was used as endogenous control. Bars represent expression in HIF-MSCs relative to GFP-MSC controls. Mean ± SD of six to eight samples pooled from at least three independent experiments is shown. Student t test, # p ≤ 0.05, ## p ≤ 0.005, GFP-MSCs versus HIF-MSCs. IL interleukin

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