Journal: Blood

Article Title: VEGFR1 stimulates a CXCR4-dependent translocation of megakaryocytes to the vascular niche, enhancing platelet production in mice.

doi: 10.1182/blood-2011-09-378174

Figure Lengend Snippet: Figure 1. VEGFR1 stimulation, but not VEGFR2 stimu- lation, is responsible VEGF-A165–induced thrombocy- tosis. (A-B) Mice were administered either VEGF-A165 (100 g/kg intraperitoneally), PlGF-2, VEGF-E (100 g/kg intraperitoneally), or vehicle (PBS) each day for 4 con- secutive days. (A) Some groups of mice receiving the 4-day VEGF-A165 regimen were also administered block- ing Abs to either VEGFR1, VEGFR2, VEGFR1&2, or control IgG 20 minutes before VEGF-A165 administration on days 1 and 3. Enumeration of circulating platelets was then conducted 24 hours after last growth factor adminis- tration. Bone marrow or spleens were also harvested, and MKs were identified by flow cytometry via forward and side scatter and CD41 expression. (C) Percentage of MKs expressing VEGFR1 and VEGFR2 in vehicle con- trol–treated mice. (D) The number of MKs per million cells from bone marrow or spleen; n 5-6 animals per group. Data are expressed as mean SEM; *P .05; ***P .001 compared with control or as indicated.

Article Snippet: In some experiments, VEGF-A165–treated mice were administered previously defined neutralizing doses of either anti-VEGFR1 Ab (50 g/mouse IP; R&D Systems), anti-VEGFR2 Ab (50 g/mouse IP; R&D Systems), or both, 2 hours before VEGF-A165 administration on days 0 and 2.9,19 Chromopure goat IgG (Jackson ImmunoResearch Laboratories) was administered to control groups.

Techniques: Blocking Assay, Control, Cytometry, Expressing

Journal: Blood

Article Title: VEGFR1 stimulates a CXCR4-dependent translocation of megakaryocytes to the vascular niche, enhancing platelet production in mice.

doi: 10.1182/blood-2011-09-378174

Figure Lengend Snippet: Figure 2. Effect of VEGFR1 and VEGFR2 stimulation on MK maturation. Mice were administered either VEGF-A165 (100 g/kg intraperitoneally), PlGF-2, VEGF-E (100 g/kg intraperitoneally), or vehicle (PBS) each day for 4 consecutive days. (A-B) Representative histograms of CD41 MKs stained with TOPRO-3 revealing respective peaks of increasing DNA ploidy number. (A) CD41 MKs with 2n, and (B) 2n peak subtracted to reveal 4n to 128n. (C) Percentage of bone marrow and splenic MKs with a ploidy number of 16n or greater in mice administered either VEGF-A165, PlGF-2, or VEGF-E. (D) Histogram revealing spread of bone marrow MK ploidy above 16n in mice administered either VEGF-A165, PlGF-2, or VEGF-E; n 5-6 animals per group. Data are expressed as mean SEM; *P .05 com- pared with control.

Article Snippet: In some experiments, VEGF-A165–treated mice were administered previously defined neutralizing doses of either anti-VEGFR1 Ab (50 g/mouse IP; R&D Systems), anti-VEGFR2 Ab (50 g/mouse IP; R&D Systems), or both, 2 hours before VEGF-A165 administration on days 0 and 2.9,19 Chromopure goat IgG (Jackson ImmunoResearch Laboratories) was administered to control groups.

Techniques: Staining, Control

Journal: Blood

Article Title: VEGFR1 stimulates a CXCR4-dependent translocation of megakaryocytes to the vascular niche, enhancing platelet production in mice.

doi: 10.1182/blood-2011-09-378174

Figure Lengend Snippet: Figure 5. VEGFR1 increases MK migration to vascu- lar niche via a CXCR4-dependent process. Mice were administered either VEGF-A165, PlGF-2, VEGF-E (100 g/kg intraperitoneally), or vehicle (PBS) each day for 4 consecutive days before bone marrow and spleens were harvested 24 hours later. (A) Percentage of CD41

Article Snippet: In some experiments, VEGF-A165–treated mice were administered previously defined neutralizing doses of either anti-VEGFR1 Ab (50 g/mouse IP; R&D Systems), anti-VEGFR2 Ab (50 g/mouse IP; R&D Systems), or both, 2 hours before VEGF-A165 administration on days 0 and 2.9,19 Chromopure goat IgG (Jackson ImmunoResearch Laboratories) was administered to control groups.

Techniques: Migration

Journal: Blood

Article Title: VEGFR1 stimulates a CXCR4-dependent translocation of megakaryocytes to the vascular niche, enhancing platelet production in mice.

doi: 10.1182/blood-2011-09-378174

Figure Lengend Snippet: Figure 4. VEGFR1 stimulation redistributes MKs to the vascular niche. Mice were administered either VEGF-A165, PlGF-2, VEGF-E (100 g/kg intraperitoneally), or vehicle (PBS) each day for 4 consecutive days before femurs were excised for bone marrow histology. (A) Quantification of MKs per field of view, and (B) quantification of the number of MKs adjacent to sinusoidal vessels; n 5-6 animals per group. Data are expressed as mean SEM; ***P .001 compared with control.

Article Snippet: In some experiments, VEGF-A165–treated mice were administered previously defined neutralizing doses of either anti-VEGFR1 Ab (50 g/mouse IP; R&D Systems), anti-VEGFR2 Ab (50 g/mouse IP; R&D Systems), or both, 2 hours before VEGF-A165 administration on days 0 and 2.9,19 Chromopure goat IgG (Jackson ImmunoResearch Laboratories) was administered to control groups.

Techniques: Control