Figure 4
Endothelial cells are effectively targeted in Tie2-Cre-loxP-GFP reporter mice and are the main source of G-csf after in vivo LPS stimulation. (A) Graphical scheme depicting experimental outline to induce LPS-induced emergency granulopoiesis and to assess G-csf expression in sorted CD45−Ter119−GFP+CD31+ ECs and CD45−Ter119−GFP+CD31− non-ECs isolated from different organs of Tie2-Cre-loxP-GFP reporter mice. (B) Representative FACS profile of GFP and CD31 expression within nonhematopoietic cells (CD45−Ter119−) isolated from BM, heart, liver, kidney, spleen, and lung of Tie2-Cre-loxP-GFP reporter mice. (C) Percentages of CD31+ (black bars) and CD31− (white bars) cells within the CD45−Ter119−GFP+ cell population in BM, heart, liver, kidney, spleen, and lung of Tie2-Cre-loxP-GFP reporter mice. (D) Percentages of CD31+ (black bars) and CD31− (white bars) cells within the CD45−Ter119−GFP− cell population in BM, heart, liver, kidney, spleen, and lung of Tie2-Cre-loxP-GFP reporter mice. (E) Comparative G-csf transcript levels normalized to Actb in sorted CD45−Ter119−GFP+CD31+ ECs (red bars) vs CD45−Ter119−GFP+CD31− non-ECs (white bars) isolated from BM, heart, liver, kidney, spleen, and lung of LPS-injected Tie2-Cre-loxP-GFP reporter mice. All data represent mean ± standard deviation from 2 independent experiments (n = 5 mice). Two-tailed Student t tests were used to assess statistical significance (*P < .05, **P < .01, ***P < .001).

Endothelial cells are effectively targeted in Tie2-Cre-loxP-GFP reporter mice and are the main source of G-csf after in vivo LPS stimulation. (A) Graphical scheme depicting experimental outline to induce LPS-induced emergency granulopoiesis and to assess G-csf expression in sorted CD45Ter119GFP+CD31+ ECs and CD45Ter119GFP+CD31 non-ECs isolated from different organs of Tie2-Cre-loxP-GFP reporter mice. (B) Representative FACS profile of GFP and CD31 expression within nonhematopoietic cells (CD45Ter119) isolated from BM, heart, liver, kidney, spleen, and lung of Tie2-Cre-loxP-GFP reporter mice. (C) Percentages of CD31+ (black bars) and CD31 (white bars) cells within the CD45Ter119GFP+ cell population in BM, heart, liver, kidney, spleen, and lung of Tie2-Cre-loxP-GFP reporter mice. (D) Percentages of CD31+ (black bars) and CD31 (white bars) cells within the CD45Ter119GFP cell population in BM, heart, liver, kidney, spleen, and lung of Tie2-Cre-loxP-GFP reporter mice. (E) Comparative G-csf transcript levels normalized to Actb in sorted CD45Ter119GFP+CD31+ ECs (red bars) vs CD45Ter119GFP+CD31 non-ECs (white bars) isolated from BM, heart, liver, kidney, spleen, and lung of LPS-injected Tie2-Cre-loxP-GFP reporter mice. All data represent mean ± standard deviation from 2 independent experiments (n = 5 mice). Two-tailed Student t tests were used to assess statistical significance (*P < .05, **P < .01, ***P < .001).

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