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Cells were fixed with 4% paraformaldehyde in PBS for 30 min and washed with PBS Ca 2+ /Mg 2+ (Sigma-Aldrich)

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One potential explanation for the significantly increased chimerism of the secondary transplants of the young Mda5 -/- is the increase in the frequency of the LSKs and HSCs in the primary transplant (not significant due to the transplanted animals with very low chimerism)