Figure 2. Flow cytometric analysis.

(A–D) Gating scheme used to identify mouse hematopoietic stem cells which were defined as side population (A), lineage negative (B), c-Kit⁺Sca-1⁺ (C). This population was then gated to a Sca-1/GFP plot to identify the proportion of HSCs that were GFP⁺. (D) HSCs from TET-GFP mice prior to administration of doxycycline were 95% GFP⁺. (E–F) Proportion of GFP⁺ cells in whole bone marrow of TET-GFP mice prior to dox treatment (E) and after 12 weeks of dox treatment (F). The percentage of GFP⁺ cells in whole bone marrow decreased in response to dox. (G) HSCs from TET-GFP mice after 12 weeks treatment with dox were still over 85% GFP⁺. (H) However, after 12 weeks of dox treatment not all GFP⁺ cells from TET-GFP mice were HSCs as only 5% of GFP⁺Lin⁻ cells were Sca-1⁺c-Kit⁺. The percentage of GFP⁺ whole bone marrow cells from TET-GFP mice decreased in response to dox (J) while the majority of HSCs remained GFP⁺ (K).