Transfection of miR155-5p mimic and inhibitor in normal and PMF CD34⁺ cells. (A) Effect of miR155-5p mimic transfection on normal CD34⁺ cell differentiation. (i) Expression levels of JARID2 at 24 and 48 hours after the last nucleofection, measured by qRT-PCR and reported as RQ. (ii-iii) Results of statistical analysis on the percentage of CD41⁺ cells performed by flow cytometry at days 5, 8, 10, and 12 after the last nucleofection on serum-free multilineage and MK unilineage cultures. (iv) Results of the statistical analysis of the collagen-based clonogenic assay. The cells were plated 24 hours after the last nucleofection and scored after 12 days. (v) Morphologic analysis of Neg-mimic and miR-155-5p nucleofected cells after MGG staining at days 8 and 10 of MK unilineage serum-free liquid culture after the last nucleofection in a representative experiment. Magnification ×1000. Values are reported as mean ± SEM. **P < .01 vs Neg-mimic; *P < .05 vs Neg-mimic. The results come from 5 independent experiments. (B) Effect of miR155-5p downregulation in PMF CD34⁺ cells. (i) Expression levels of JARID2 in PMF CD34⁺ cells at 24 and 48 hours after the last nucleofection of miR-155-5p inhibitor. The JARID2 expression was measured by qRT-PCR, and data are reported as RQ. (ii-iii) Percentage of viable CD41⁺ cells assessed by flow cytometry at days 5, 8, and 10 after the last nucleofection on serum-free multilineage and MK unilineage cultures. (iv) Results of the statistical analysis of the collagen-based clonogenic assay. The cells were plated 24 hours after the last nucleofection and scored after 12 days. (v) Morphologic analysis of negative control inhibitor (NegINH) and miR-155-5p inhibitor treated cells after MGG staining at days 8 and 10 of MK unilineage serum-free liquid culture after the last nucleofection in a representative experiment. Magnification ×1000. Values are reported as mean ± SEM. **P < .01 vs NegINH; *P < .05 vs NegINH. The results come from 4 independent experiments.