Fig. 2. VMP1 knockdown in HSPCs results in inhibition of autophagy and an impaired in vitro expansion and in vivo engraftment.

a Experimental scheme, shSCR-mCherry or shVMP1-mCherry transduced cord blood (CB) CD34⁺ cells were cultured in vitro under myeloid or erythroid liquid culture conditions or injected IV in sub-lethally irradiated NSG mice. Bleeds were performed at week 5, 8, and 12 after injection and analysed by FACS. b Knockdown efficiency of VMP1 was determined by quantitative RT–PCR and Western blotting of CB CD34⁺ cells transduced with shSCR, shVMP1 #1 or shVMP1 #2. c CFC assay with freshly sorted shSCR or shVMP1 #1 transduced CB CD34⁺ cells (n = 2). d Representative graph showing relative expansion of shVMP1 transduced CB CD34⁺ cells under myeloid or erythroid permissive liquid culture conditions, shSCR transduced cells were used as control (n = 2). e Percentage of engraftment represented by huCD45 percentage (left graph) and the percentage mCherry within the huCD45⁺ population (right graph). Each dot represents data from a single mouse, n = 5 for each group. Error bars represent SD; *, ** or *** represents p < .05, p < .01 or p < .001, respectively