Fig. 3. Alterations in hematopoietic cells and defective emergency hematopoiesis in Mlkl^D139V mice.

a–c Absolute white blood cell (WBCB), lymphocyte and platelet numbers in peripheral blood of E19.5 and P3 pups, n = 6, 27, 44, 41, 10, and 11 as indicated. d Proportions of HSC (Lineage^-Sca-1⁺c-kit⁺ (LSK) CD150⁺ CD48⁻), MPP (LSK CD150⁻ CD48⁻), HPC-1 (LSK CD150⁻ CD48⁺) and HPC-2 (LSK CD150⁺ CD48⁺)⁸², n = 5 per genotype and (e) relative levels of ROS (n = 4, 9, 5) (f) P2 bone marrow LSK populations (n = 9, 18, and 11) and (g) relative AnnexinV binding (n = 2, 11, 7). (h) HSC subtypes in adult bone marrow, n = 9 per genotype. a–h Each symbol represents one independent animal; Mlkl^Wt/Wt – blue circles, Mlkl^Wt/D139V- red squares, Mlkl^D139V/D139V- green triangles, with bar height and error bars representing mean ± SD respectively, or range when n = 2. Red and white blood cells and platelets in Mlkl^Wt/Wt (blue circles) and Mlkl^Wt/D139V (red squares) mice after treatment with 150 mg per kg 5FU or saline. Means ± SEM from one experiment in which three mice were sampled at each time point for each treatment group, similar results were obtained in an independent cohort. j Bone marrow from Mlkl^Wt/Wt or Mlkl^Wt/D139V mice on CD45^Ly5.2 background was mixed with wild-type CD45^Ly5.1 competitor bone marrow and transplanted into irradiated CD45^Ly5.1/Ly5.2 recipients. Peripheral blood mononuclear cells (PBMCs) quantified after 56 and 180 days. Mean ± SEM are shown (3 donors per genotype, 3–5 recipients per donor). k Fetal liver cells (CD45^Ly5.2; Mlkl^Wt/Wt, Mlkl^Wt/D139V or Mlk^D139V/D139V) were transplanted into lethally irradiated recipients (CD45^Ly5.1/Ly5.2) together with competitor bone marrow (CD45^Ly5.1). Contribution to PBMCs 28 days and 180 days after transplantation. Mean ± SEM are shown (2–10 donors per genotype, 2–6 recipients per donor). Host contribution (CD45^Ly5.1/Ly5.2) is depicted in gray, competitor (CD45^Ly5.1) in white, and test (CD45^Ly5.2) in black for (j) and (k). *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.005 calculated using an unpaired, two-tailed t-test.