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.