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. 2022 Nov 1;132(21):e158869. doi: 10.1172/JCI158869

Figure 1. Samd9l-Mut mouse has altered hematopoiesis.

Figure 1

(A) Model for the generation of conditional knockin Samd9l-W1171R mutation (Samd9l cKI+/+) crossed with the hematopoietic cell–specific Vav1-Cre mouse (Vav1-Cre+/Tg) to produce offspring with heterozygous Samd9l mutations (Vav1-Cre+/Tg Samd9l cKI+/–). (B) PCR analysis verifying the genotypes of the mice. The left gel shows the PCR result for the knockin insertion to have a 1,270-bp product if the cassette is present, 514-bp product if not, and both for heterozygosity. The right gel shows the PCR products for the Vav-1 amplicon using iCre primers (390 bp) and internal positive control (324 bp). (C) Complete blood count (CBC) of Samd9l-KO (Samd9l–/–, blue, n = 6), Samd9l-WT (Samd9lcKI+/–, black, n = 11), and Samd9l-Mut (Vav1-Cre+/Tg Samd9lcKI+/–, red, n = 14) mice at 3 months: white blood cells (WBC, left), red blood cells (RBC, middle), and platelets (PLT, right). (D and E) Flow cytometric analysis of C57BL/6 (gray, n = 4), Samd9l-KO (blue, n = 8), Samd9l-WT (black, n = 8), and Samd9l-Mut (red, n = 8) mice assessing the BM compartment for (D) Lineage– (Lin–), common lymphoid progenitors (CLPs), myeloid progenitors (MPs), common myeloid progenitors (CMPs), granulocyte-macrophage progenitors (GMPs), and megakaryocyte/erythroid progenitors (MEPs), and (E) KSL (Lin–cKit+Sca-1+), multipotent progenitors (MPP2 and MPP3/4), and short-term and long-term HSCs (ST-HSCs and LT-HSCs). (F and G) Percentage of mature cells in (F) peripheral blood (PB) or (G) BM cells of the C57BL/6, Samd9l-KO, Samd9l-WT, and Samd9l-Mut mice assessed by flow cytometry. For panels C–G, groups were initially compared by Kruskal-Wallis test. Significant Kruskal-Wallis results were followed by pairwise comparisons with Wilcoxon’s rank-sum test. *P < 0.05; **P < 0.01; ***P < 0.001. Error bars indicate the SEM for biological replicates.