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. 2013 Mar 11;210(3):623–639. doi: 10.1084/jem.20121610

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© 2013 Brenet et al.

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Figure 4. — Blockade of TGFβ during recovery from myelotoxic stress prolongs HSPC cycling. (A) Flow cytometry gating strategy for cell cycle analysis of LKS⁺ (Lin⁻cKit⁺Sca1⁺) and LKS + SLAM (Lin⁻cKit⁺Sca1⁺CD48⁻CD150⁺) cells is shown (top images show frequency in total BM; bottom images show percentage of LKS + SLAM gated cells). (B) Mice were treated with 5FU, and then with the TGFβ-neutralizing antibody, 1D11 (5FU-I), or the isotype control antibody 13C4 (5FU-C) on day 5, 7, and 9. The percentage of quiescent LKS⁺ cells is shown before treatment (D0) and at various times after treatment with 5FU and 1D11 (5FU-I, gray bars) or the control antibody (5FU-C, white bars; n = 5/group). (C) Cell cycle status of BM LKS + SLAM HSCs is shown before (D0) and on day 15 after treatment with 5FU and 1D11 (5FU-I) or the control antibody (n = 5). (D) Cohorts of untreated, homeostatic mice (n = 10) were administered either the TGFβ-neutralizing antibody 1D11 (I; 10 mg/kg) or control antibody 13C4 (C; 10 mg/kg) on days −3 and −1 before analysis on day 0. Bivariate cell cycle status of BM LKS + SLAM HSC on day 0 is shown. All quantified data are shown as mean ± SEM (*, P < 0.05; **, P < 0.01; ***, P < 0.001, or if undesignated, the comparison was not significant).