Fig. 3

PRLR signaling modulates the in vivo AML regeneration capacity. A rFluc-transduced MonoMac-1 (MM) were intravenously injected into adult conditioned NSG mice (n = 34). At day 4, mice were treated 5 days with the vehicle (grey), PRL (blue) or G129R (red) at 0.2 mg/kg. Engraftment was detected by bioluminescence. A representative bioluminescence image at day 13 is presented. *p < 0.05; **p < 0.01; ****p < 0.0001 (one-way ANOVA, Tukey’s multiple comparison test). B Activity of the Stat5-reporter CISH was evaluated in the presence of the empty vector (grey), PRL wt (orange) or PRL mut (blue) PRLR-expressing HEK293T cells. ***p < 0.001 (one-way ANOVA, Tukey’s multiple comparison test, triplicates). C Parental (grey) and PRL wt- (orange) or PRL mut- (blue) transduced MonoMac-1 (MM, n = 7) and HL-60 (n = 5) were cultured in methylcellulose and colonies were counted after 7 days. *p < 0.05; **p < 0.01 (non-parametric one-way ANOVA, Tukey’s multiple comparison test). D Activation of Stat5 reporter CISH was assessed in PRLR wt- (orange) and PRLR short- (blue) transfected HEK293T cells treated with increasing doses of PRL. *p < 0.05; **p < 0.01 (one-way ANOVA, Šidák’s multiple comparison test, triplicates). E An equivalent number of pSmurf-transduced MonoMac-1 (blue), and empty control or PRL wt/PRL mut (n = 28) or PRLR wt/PRLR short (n = 31) MonoMac-1 cells (yellow) were intravenously injected into adult conditioned NSG mice. After 15 days, CD45⁺ AML cells from bone marrow and spleen were analysed by flow cytometry. *p < 0.05; ***p < 0.001; ****p < 0.0001 (non-parametric one-way ANOVA, Tukey’s multiple comparison test). F Migration capacity was determined using a transwell chamber assays. Growth medium containing 10% FBS was placed in the lower chamber and MonoMac-1 (MM) cells were incubated with the vehicle (grey) or PRL at 500 ng/mL (blue). Cells were allowed to migrate into the low chamber for 48 h. Migrated cells were quantified by flow cytometry. *p < 0.05 (unpaired t test, triplicates). In all graphs, bars represent the mean ± SEM