Effects of hsa-miR-223 or hsa-miR-125b overexpression on protein and mRNA levels of genes with roles in B-cell maturation. (A) Effect of the overexpression of hsa-miR-223 on native LMO2 protein levels in VAL cell line at 72 hours after transfection, assessed by Western blot. (B) PRDM1 and IRF4 protein levels detected by Western blot in RCK8 48 hours after transfection of hsa-miR-125b precursor. GAPDH levels were used as loading control in all cases. Data were confirmed in duplicate experiments. Effects of the overexpression of hsa-miR-223 on the mRNA levels of LMO2 (C), or of the overexpression of hsa-miR-125b on mRNA levels of PRDM1 (D left upper panel) and IRF4 (D left lower panel), measured by real-time PCR using TaqMan Gene Expression Assays (Applied Biosystems) at 24, 48, and 72 hours after transfection. Values of triplicate wells are represented as fold expression with respect to the nontargeting control transfection. Overexpression of both miRNAs was confirmed by TaqMan MicroRNA Assays (C right panel and D right panel), expressed as fold increase regarding the control transfection. Error bars correspond to standard error of the mean in all graphs.