Figure 2
Figure 2. Activation of PTH signaling in osteocytes does not change BM cellularity, phenotypic HSCs, spleen weight, or extramedullary hematopoiesis. (A-C) Representative flow cytometric dot plot illustrating gating scheme for subsets of LSK cells from the BM from 3.5- to 4-week-old WT or TG male littermate mice. Parent population is obtained from the FSC and SSC population. The lineage-negative, DAPI-negative population (A) was further analyzed for Sca-1 and c-kit expression (B).The LSK cell subset was further analyzed for the expression of the SLAM receptors CD150 and CD48 (representative contour plot in panel C) and for Flt-3 (not shown). (D) Total BMMCs obtained from crushing 1 femur and 1 tibia from each mouse are depicted. LSK cells (E) and subsets for MPPs/ST-LSK (Flt3+CD48−CD150−, F) and LT-LSK (Flt3−CD48−CD150+, G) are represented as frequency of total. (H) Spleen weights in 4-week-old WT or TG male littermates. (I-K) Spleen LSK cells (I), ST-HSCs (J), and LT-HSCs (K). The analysis was performed in 2 separate experiments for the BM (WT, n = 7; TG, n = 9), 4 separate experiments for the spleen (WT, n = 17; TG, n = 18; spleen weights were obtained in 2 of 4 experiments). Each dot represents an individual mouse; each separate experiment is indicated by a different color; bar indicates the mean and SEM. None of the results demonstrated a statistically significant difference.

Activation of PTH signaling in osteocytes does not change BM cellularity, phenotypic HSCs, spleen weight, or extramedullary hematopoiesis. (A-C) Representative flow cytometric dot plot illustrating gating scheme for subsets of LSK cells from the BM from 3.5- to 4-week-old WT or TG male littermate mice. Parent population is obtained from the FSC and SSC population. The lineage-negative, DAPI-negative population (A) was further analyzed for Sca-1 and c-kit expression (B).The LSK cell subset was further analyzed for the expression of the SLAM receptors CD150 and CD48 (representative contour plot in panel C) and for Flt-3 (not shown). (D) Total BMMCs obtained from crushing 1 femur and 1 tibia from each mouse are depicted. LSK cells (E) and subsets for MPPs/ST-LSK (Flt3+CD48CD150, F) and LT-LSK (Flt3CD48CD150+, G) are represented as frequency of total. (H) Spleen weights in 4-week-old WT or TG male littermates. (I-K) Spleen LSK cells (I), ST-HSCs (J), and LT-HSCs (K). The analysis was performed in 2 separate experiments for the BM (WT, n = 7; TG, n = 9), 4 separate experiments for the spleen (WT, n = 17; TG, n = 18; spleen weights were obtained in 2 of 4 experiments). Each dot represents an individual mouse; each separate experiment is indicated by a different color; bar indicates the mean and SEM. None of the results demonstrated a statistically significant difference.

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