Journal: The Journal of Experimental Medicine
Article Title: Hematopoietic Stem Cells Are Uniquely Selective in Their Migratory Response to Chemokines
doi: 10.1084/jem.20011284
Figure Lengend Snippet: LT-HSC and ST-HSC contain mRNA for CCR3, CCR9, and CXCR4. (A) RT-PCR analysis of mRNA for chemokine receptors of combined LT-HSC and ST-HSC (Thy-1.1 lo Sca-1 + Lin −/lo c-Kit + cells). RT-PCR was performed on RNA isolated from the equivalent of 1,000 double-sorted HSC, or from 1,000 unfractionated WBM cells. Representative data are shown (see for data summary). (B) Additional RT-PCR was performed on mRNA isolated from the equivalent of 1,000 LT-HSC or 1,000 ST-HSC for the receptors found to be positive in the first screen. Both LT-HSC and ST-HSC contained mRNA for CXCR4, CCR3, and CCR9. See the Materials and Methods section for RT-PCR protocol. DHFR, dihydrofolate reductase; WBM, whole bone marrow.
Article Snippet: The following chemokines were added to the bottom well, and cells were allowed to migrate for 2 h at 37°C: mouse JE, mouse eotaxin, human thymus- and activation-regulated chemokine, mouse regulated upon activation, normal T expressed and secreted (RANTES), mouse macrophage inflammatory protein (MIP)-1α, human MIP-3α, human I-309, mouse KC, and human SDF-1α (PeproTech); mouse MIP-1β, human MIP-3β, mouse thymus–expressed chemokine (TECK), mouse monokine induced by IFN-γ (MIG), and mouse B lymphocyte chemokine (BLC)/BCA-1 (R&D Systems); and human IL-8 (a gift from K. Matsushima, University of Tokyo, School of Medicine, Tokyo, Japan).
Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation