TSC2 deletion in myeloid cells impairs medullary erythropoiesis. (A) Schematic representation of the genetic background of cre/+ mice vs +/+ controls. TSC2 knockout mice are generated by LysZ-cre system cross-targeting TSC2 gene with loxP sites (cre/+ mice). Control littermates contain the TSC2 gene flanked by LoxP sites (+/+). (B) Representative image showing defect in medullary erythropoiesis in the BM isolated from control (+/+) and cre/+ mice. (C) Bar graph of flow cytometric analysis showing erythroblast counts in the BM (∗P < .05; ∗∗P < .01). Different maturation stages of erythroblasts (pro-Erys, proerythroblasts; I, basophilic erythroblasts; II, orthochromatic erythroblasts; III, polychromatic erythroblasts; and IV, acidophilic erythroblasts) were identified based on CD71 (Tf receptor) and Ter119 (erythrocyte marker) expression and cell size (SSC). (D) Photography of the spleen of cre/+ mice compared with controls (+/+) represents splenomegaly in cre/+ mice. (E) Bar graph of flow cytometric analysis of erythroblast counts in different maturation stages in the spleen identified via CD71 (Tf receptor) and Ter119 (erythrocyte marker) expression (∗P < .05; ∗∗∗P < .001). (F-K) Complete blood cell count analysis showing hematological parameters in whole blood from control (+/+) and cre/+ mice: RBC count (F), blood hemoglobin (HGB) level (G), mean corpuscular volume (MCV) of RBCs (∗∗∗∗P < .0001) (H), mean corpuscular hemoglobin (MCH) of RBCs (∗∗∗∗P < .0001) (I), mean corpuscular hemoglobin concentration of RBCs (∗P < .05) (J), and the red cell distribution width (RDW) of RBCs (∗∗∗∗P < .0001) (K).