Figure 7

Deletion of THOC5/FMIP causes loss of primitive hematopoietic cells from the bone marrow. Five-to-six-week-old Mx-cre THOC5/FMIP (flox/flox) (n = 5) and THOC5/FMIP (flox/flox) (n = 6) mice were injected with poly (I:C) (2 × 500 μg, two days interval). Then, bone marrow cells were isolated from mice four days after poly (I:C) injection. The effect of THOC5/FMIP depletion on the total numbers of primitive cells present in the bone marrow was assessed using a number of different approaches (A; B): total bone marrow cellularity per femur was assessed (A) and the number of cells present in the bone marrow which do not express lineage markers (B) (and therefore have a primitive phenotype) was calculated (Mean+/-SEM). Results shown are for Mx-cre THOC5/FMIP (flox/flox) (Mx-CreFMIPf/f) mice with (n = 5) and without (n = 4) poly (I:C) (2 × 500 μg, two day interval), and THOC5/FMIP (flox/flox) (FMIPf/f) control mice with (n = 6) and without (n = 6) Poly (I:C) (2 × 500 μg, two day interval) (C). The number of colony forming cells per femur was determined for Colony Forming Unit-Granulocyte macrophage (GM-CFU) and Colony Forming Unit-Granulocyte Erythroid Macrophage Megakaryocyte (GEMM-CFU). Results shown are the mean+/-SEM, n = 3. (D): Flow cytometric profile of Kit and Sca staining in Lineage marker depleted cells from mice treated with and without poly (I:C) (3 × 250 μg, two day interval). The results shown are representative of six experiments. (E; F) Results shown are for Mx-cre THOC5/FMIP (flox/flox) mice with (n = 5) and without (n = 4) poly (I:C) (2 × 500 μg, two day interval), and THOC5/FMIP (flox/flox) control mice with (n = 6) and without (n = 6) poly (I:C) (2 × 500 μg, two-day-interval), and are expressed as total number of LSK cells (E) and Lin-Sca-Kit+ cells (LS-K+) cells (F) per femur, error bars show SEM.