HIF-1⍺ promotes MerTK cleavage through glycolytic reprogramming of macrophages. (A and B) Cell surface expression of MerTK (A) or solMER (B) in culture media of untreated (⍉) or LPS-treated BMDMs from controls or mice with myeloid-specific deletion of Hif2 (mHIF2−/−). (C) Efferocytosis of calcein-labeled apoptotic Jurkat cells (green) by BMDMs from mice with myeloid-specific deletion of Hif1 (mHIF1−/−) or controls treated with LPS. Scale bar, 10 µm. n = 3 sets of cells/group. Data are representative of two independent experiments. *, P < 0.05 by two-way ANOVA followed by Tukey’s test. (D and E) Cell surface expression of MerTK (D) or solMER (E) in culture media of untreated (⍉) or LPS-treated BMDMs from controls, mHIF1−/− mice, or mice with overexpression of HIF-1⍺ (mHIF1LSL). For B–E, n = 3–4 sets of cells/group, and data are representative of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. (F) Expression of Adam17 in untreated (⍉) or LPS-treated mHIF1−/− BMDMs or controls. n = 3 sets of cells/group and data are representative of three independent experiments. *, P < 0.05 by two-way ANOVA followed by Tukey’s test. (G) Expression of Adam17 in untreated mHIF1LSL BMDMs or controls. n = 3 sets of cells/group, and data are representative of two independent experiments. *, P < 0.05 by two-tailed, unpaired t test. (H) ECAR with quantification of glycolytic function in untreated (⍉) or LPS-treated BMDMs. n = 5–7 sets of cells/group. Data are representative of more than three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. (I and J) Cell surface expression of MerTK (I) or solMER (J) in culture media of untreated or LPS-treated BMDMs cultured in 0 mM glucose (Glu), 5 mM glucose, or 5 mM glucose with 25 mM 2-DG. n = 4 sets of cells/group. Data are representative of two independent experiments. **, P < 0.01; ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. Unfilled histograms represent FMO staining controls. All data presented as mean ± SEM.
Figure 5.

HIF-1⍺ promotes MerTK cleavage through glycolytic reprogramming of macrophages. (A and B) Cell surface expression of MerTK (A) or solMER (B) in culture media of untreated (⍉) or LPS-treated BMDMs from controls or mice with myeloid-specific deletion of Hif2 (mHIF2−/−). (C) Efferocytosis of calcein-labeled apoptotic Jurkat cells (green) by BMDMs from mice with myeloid-specific deletion of Hif1 (mHIF1−/−) or controls treated with LPS. Scale bar, 10 µm. n = 3 sets of cells/group. Data are representative of two independent experiments. *, P < 0.05 by two-way ANOVA followed by Tukey’s test. (D and E) Cell surface expression of MerTK (D) or solMER (E) in culture media of untreated (⍉) or LPS-treated BMDMs from controls, mHIF1−/− mice, or mice with overexpression of HIF-1⍺ (mHIF1LSL). For B–E, n = 3–4 sets of cells/group, and data are representative of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. (F) Expression of Adam17 in untreated (⍉) or LPS-treated mHIF1−/− BMDMs or controls. n = 3 sets of cells/group and data are representative of three independent experiments. *, P < 0.05 by two-way ANOVA followed by Tukey’s test. (G) Expression of Adam17 in untreated mHIF1LSL BMDMs or controls. n = 3 sets of cells/group, and data are representative of two independent experiments. *, P < 0.05 by two-tailed, unpaired t test. (H) ECAR with quantification of glycolytic function in untreated (⍉) or LPS-treated BMDMs. n = 5–7 sets of cells/group. Data are representative of more than three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. (I and J) Cell surface expression of MerTK (I) or solMER (J) in culture media of untreated or LPS-treated BMDMs cultured in 0 mM glucose (Glu), 5 mM glucose, or 5 mM glucose with 25 mM 2-DG. n = 4 sets of cells/group. Data are representative of two independent experiments. **, P < 0.01; ***, P < 0.001 by two-way ANOVA followed by Tukey’s test. Unfilled histograms represent FMO staining controls. All data presented as mean ± SEM.

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