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. Author manuscript; available in PMC: 2021 May 1.
Published in final edited form as: Leukemia. 2020 Jun 24;34(11):2951–2963. doi: 10.1038/s41375-020-0929-3

Figure 1. Supplementation of primary CMML cells with MN1 promotes engraftment in vivo.

Figure 1

A) Engraftment of human hematopoietic cells (CD45 positive) in the peripheral blood of primary recipient NSGS mice monitored at the indicated time points. Recipient mice were transplanted with untransduced CMML#1 cells (CMML#1-CTL), EGFP- (control vector) transduced CMML#1 cells (CMML#1-EGFP) or MN1-transduced CMML#1 cells (CMML#1-MN1), (number of analyzed mice is indicated in the figure; mean ± SEM).

B) Engraftment of human CD45 positive cells in peripheral blood, bone marrow and spleen of primary recipient NSGS mice at sacrifice (number of analyzed mice is indicated in the figure; mean ± SEM).

C) Expression levels of MN1 in untransduced CMML#1 patient cells (CMML#1-CTL), EGFP-transduced CMML cells (CMML#1-EGFP) and MN1-transduced CMML cells (CMML#1-MN1). Gene expression was determined by quantitative RT-PCR relative to the housekeeping gene ABL1 and normalized to gene expression in untransduced CMML#1 patient cells (n=3), (mean ± SEM).

D) Western blot showing MN1 expression in primary bone marrow mononuclear cells from a patient with CMML (CMML#1-CTL) and in their MN1-transduced counterparts isolated from bone marrow of engrafted recipient mice. ß-actin was used in the same blot as loading control.

E) Spleen weight of primary recipient NSGS mice at sacrifice (number of analyzed mice is indicated in the figure; mean ± SEM).

F) Platelet count in peripheral blood of primary recipient NSGS mice at sacrifice (number of analyzed mice is indicated in the figure; mean ± SEM).

G) White blood cell count in peripheral blood of primary recipient NSGS mice at sacrifice (number of analyzed mice is indicated in the figure; mean ± SEM).

H) Hemoglobin level in peripheral blood of primary recipient NSGS mice at sacrifice (number of analyzed mice is indicated in the figure; mean ± SEM).

I) Immunophenotype of engrafted CMML#1 cells from bone marrow and spleen of primary recipient NSGS mice at sacrifice (gated on human CD45+EGFP+ CMML#1-MN1 cells; n=3; mean ± SEM).

J) Morphology from bone marrow smears of the CMML#1 patient at diagnosis and from a transplanted mouse at sacrifice. The enlarged cell shows granules in the cytoplasm that can be identified as lipid granules in cells from later transplantations (see Figure 3B).