![STAT5 loss leads to defective HSC function. (A) Bar plot showing the frequency of ESLAM HSCs (CD45+CD150+CD48−EPCR+) in BMMNCs from WT and STAT5-deficient mice (mean ± standard error of the mean [SEM]). (B) Bar plot showing the frequency of LT-HSCs (Lin−Sca1−cKit+CD150+CD48−CD34−Flk2−) in BMMNCs (mean ± SEM). (C) Bar plot showing the frequency of B cells (B220+) in BMMNCs (mean ± SEM). (D) Bar plot showing the frequency of CFU-e progenitors (Lin−Sca1−cKit+CD41−CD16/32−CD105+CD150−) in BMMNCs (mean ± SEM). (E) Bar plots showing the frequency of CFU-e and pre–CFU-e (Lin−Sca1−cKit+CD41−CD16/32−CD105+CD150+) cells in spleen mononuclear cells (mean ± SEM). (F) Bar plots (left) showing the frequency of megakaryocyte (CD41+CD42+) and erythroid precursor cells (I, CD71hiTer119mid; II, CD71hiTer119hi; III, CD71midTer119hi; IV, CD71lowTer119hi) in spleen mononuclear cells (mean ± SEM) with a representative flow-cytometry plot (right) showing the gating of different stages of erythroid precursor cells in terminal differentiation. (G) Schematic diagram showing that 33 fluorescence-activated cell sorting–purified BM ESLAM HSCs were transplanted into irradiated recipient mice with 5 × 105 competitor BMMNCs. STAT5 was deleted in Cre+ donor cells after transplantation by repeated injection (×7) with Poly:IC in recipients. Blood was taken before and after STAT5 deletion and was followed for 5 months after deletion before serial transplantation of 3 × 106 primary recipient BMMNCs. (H) Connected line graphs showing donor chimerism in peripheral blood mononuclear cells at each time point in primary and secondary recipients (mean ± SEM). The dotted line indicates initiation of the Poly:IC injections. The asterisks indicate significant differences by analysis of variance column factor (∗∗∗∗P < .0001). (I) Bar plots showing the total BMMNC donor chimerism in primary and secondary recipients (mean ± SEM). (J) Bar plots showing LT-HSC donor chimerism in primary and secondary recipients (mean ± SEM). The asterisks indicate significant differences by Student t test (∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05) unless otherwise indicated.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/9/2/10.1182_bloodadvances.2024014046/3/blooda_adv-2024-014046-gr1ij.jpeg?Expires=1743232483&Signature=N2dW63Ajs-tPkdGE08ZWoUToa4D0rFNOrLYnAi4d8bXa1VKJzFLtj2Mut0BNx1zQo0sUPnDLG25kXF-yyaaPQYp75-EXbyIp-wt8rMOlSTdC~iRX~QcTtxxDqE6qfVOT0v-TrGNHMBSE3Rz2YZipTzpnmsSvI2NBEepeV-toNBu38GyKT9tU~RQIY2HUwWFiegMIgVMpeCxzkWJcQiqd6hnZwbRY85WLJPmRHn~q5qMkBqdbCoP58ksv6KlWi1AJ4SeZJm4Dd43Z5Wg8QKXD6S26C7hO7YO0zGMQZ6BIwZsINrHSd41IDDQ44p6c5FJDJQL6FcjHeammPfw12~ulGA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
STAT5 loss leads to defective HSC function. (A) Bar plot showing the frequency of ESLAM HSCs (CD45+CD150+CD48−EPCR+) in BMMNCs from WT and STAT5-deficient mice (mean ± standard error of the mean [SEM]). (B) Bar plot showing the frequency of LT-HSCs (Lin−Sca1−cKit+CD150+CD48−CD34−Flk2−) in BMMNCs (mean ± SEM). (C) Bar plot showing the frequency of B cells (B220+) in BMMNCs (mean ± SEM). (D) Bar plot showing the frequency of CFU-e progenitors (Lin−Sca1−cKit+CD41−CD16/32−CD105+CD150−) in BMMNCs (mean ± SEM). (E) Bar plots showing the frequency of CFU-e and pre–CFU-e (Lin−Sca1−cKit+CD41−CD16/32−CD105+CD150+) cells in spleen mononuclear cells (mean ± SEM). (F) Bar plots (left) showing the frequency of megakaryocyte (CD41+CD42+) and erythroid precursor cells (I, CD71hiTer119mid; II, CD71hiTer119hi; III, CD71midTer119hi; IV, CD71lowTer119hi) in spleen mononuclear cells (mean ± SEM) with a representative flow-cytometry plot (right) showing the gating of different stages of erythroid precursor cells in terminal differentiation. (G) Schematic diagram showing that 33 fluorescence-activated cell sorting–purified BM ESLAM HSCs were transplanted into irradiated recipient mice with 5 × 105 competitor BMMNCs. STAT5 was deleted in Cre+ donor cells after transplantation by repeated injection (×7) with Poly:IC in recipients. Blood was taken before and after STAT5 deletion and was followed for 5 months after deletion before serial transplantation of 3 × 106 primary recipient BMMNCs. (H) Connected line graphs showing donor chimerism in peripheral blood mononuclear cells at each time point in primary and secondary recipients (mean ± SEM). The dotted line indicates initiation of the Poly:IC injections. The asterisks indicate significant differences by analysis of variance column factor (∗∗∗∗P < .0001). (I) Bar plots showing the total BMMNC donor chimerism in primary and secondary recipients (mean ± SEM). (J) Bar plots showing LT-HSC donor chimerism in primary and secondary recipients (mean ± SEM). The asterisks indicate significant differences by Student t test (∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05) unless otherwise indicated.
