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

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 Fig. 3b).