Figure 3.
GlcNAc exacerbates the severity of an advanced systemic mastocytosis (AdvSM) phenotype. (A) The proliferation of human MCs, ROSAKIT WT, and ROSAKIT D816V cells, was assessed after treatment with or without 20 mM GlcNAc, and the cells were counted using a Cellometer Auto T4 at days 0, 2, 4, and 8. (B) Schema of in vivo administration of 300 mg/kg GlcNAc to ROSAD816-Gluc transplanted mice after irradiation with 1.5 Gy (n = 10, 5 per group). (C) Measure of GlcNAc activity (in relative luciferase units [RLU]) in PBS- or GlcNAc-treated mice from week 3 (W3) to week 7 (W7). (D) Number of Gluc+ cells circulating in the plasma of mice from W3 to W7, as measured by fluorescence-activated cell sorting (FACS). (E) Number of GFP+ and CFP+ cells in the bone marrow of PBS- or GlcNAc-treated mice, as measured by FACS. (F) Number of GFP+ and CFP+ cells in the spleens of PBS- or GlcNAc-treated mice, as measured by FACS. (G) Weights of spleens at W7 in PBS or GlcNAc-treated mice. (H) Plasmatic concentration of GlcNAc measured by LC-MS/MS in nontransplanted mice, transplanted mice, and GlcNAc-treated transplanted mice. Sampling was done after 7 weeks of treatment, 2 hours after the last injection of GlcNAc. The values in the graphs are presented as the means ± SD (nonsignificant, *P < .05, **P < .01, ***P < .001; 2-tailed, unpaired Student t test).