Fig 2: Proliferation, migration, and cell stemness of BMSCs with Fgfr2 overexpression. (A) The proliferation curves of BMSCs after Fgfr2 overexpression (n = 6). (B) The transwell migration assay and quantification of BMSCs after Fgfr2 overexpression (n = 3). (C) Scratch experiment images of BMSCs after Fgfr2 overexpression (n = 3). Dashed lines represent the scratch width. (D) The flow cytometric graphs and quantification of the mean fluorescence intensity (MFI) of CD200 and CD105 after Fgfr2 overexpression (n = 3). (E) Tunel staining and quantification of BMSCs after 72 h serum shock. Graphs show the mean value ±SD. *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 3: Histograms of expression of CD44, CD90, CD105, CD34, and CD45 markers in the culture of murine ADSCs from young animals according to flow cytometry, the 2nd passage.

Fig 4: Characteristics of the LRRC15-positive cells in the mice bone marrow tissues(A) FACS analysis for the Lrrc15-expressing population in the CD11b/−CD45-/Ter119-/7-AAD- fractions.(B) Comparison of Twist1 gene expression level between the Lrrc15- and Lrrc15+ populations. Asterisk means significant difference were detected at p < 0.05 (N = 6 from different animals).(C) Colony formation properties of the FACS-sorted cells from CD11b/−CD45-/Ter119-/7-AAD-/Lrrc15- (upper) or CD11b−/CD45-/Ter119-/7-AAD-/Lrrc15+ (lower) fractions.(D) Flow cytometry analysis for the CD11b−/CD45-/Ter119-/7-AAD-/Lrrc15+ cells with the MSC markers in mice. Gray histograms show control cells treated with isotype antibodies, and pink histograms show samples treated with each antibody.(E) FACS sorting of the MSCs using anti-CD105 and anti-LRRC15 antibodies.(F) Comparing gene expressions of Twist1, stemness-associated transcription factors, and therapeutic cytokines in the CD11b−/CD45-/Ter119-/CD105+/Lrrc15- and CD11b−/CD45-/Ter119-/CD105+/Lrrc15+ cells. Asterisks mean significant differences detected at p < 0.05 (N = 3, independent experiment).(G) Engrafted EGFP+ cells were detected by FACS analysis. Numbers in the figures are mean values with S.D. from three independent animals per each group.

Fig 5: Defect in erythroid precursors in BM of Gata1s mice. (A-B) Representative flow cytometry plots (A) and bar graph (B) of mean ± SD for various stages of BM erythropoiesis in 2-month-old WT or Gata1s (G1s) male mice as assessed by staining with antibodies against Ter119 and CD44. (C-J) Analysis of the composition of BMCs by flow cytometry for hematopoietic progenitors in 2-month-old mice. Numbers indicate the percentage of cells within each gate as shown in the representative plots. (C-D) Representative flow cytometry plots and mean ± SD of MEP (Lin−c-kit+Sca-1−CD34−CD16/32−), granulocyte-monocyte progenitor (GMP) (Lin−c-kit+Sca-1−CD34+CD16/32+), and common myeloid progenitor (CMP) (Lin−c-kit+Sca-1−CD34+CD16/32−) cell populations from WT and G1s mice. (E-F) Lineage−Sca-1−c-Kit+ cells were further subdivided by expression of CD105 and CD41 into the megakaryocyte progenitors (MkPs, CD105+CD41+). (G-J) Lineage−Sca-1−c-Kit+CD34−CD16/32− cells were further subdivided by expression of CD105 and CD150 into pregranulocyte-macrophage progenitors (pre-GMs, CD105−CD150−), the premegakaryocytic/erythroid progenitors (preMegEs, CD105−CD150+), and the pre-CFU erythroid cells (preCFU-Es, CD105+CD150+). Graphs depict mean (± SD) percentage of flow cytometry data. Nwt = 3, NG1s = 4. ∗P ≤ .05, ∗∗P ≤ .01, ∗∗∗P ≤ .001 by the unpaired Student t test.