Figure 3

EPN-01 and MID-PIWI-Vpr transfection increases both the variety of miRNA species and the level of miRNAs from transfected HEK293T cells. (A) The read counts for the detected miRNAs (with a threshold of ≥ 5 read counts) obtained from two independent experiments were calculated and plotted on the x- and y-axes. (B) (left) Total number of miRNA species in the fraction of non-transfected (control) or transfected (EPN-01 + Flg-MID-PIWI-Vpr) HEK293T cells. (right) Venn diagrams of miRNAs in the EV fraction of non-transfected (control) or transfected (EPN-01 + Flag-MID-PIWI-Vpr) HEK293T cells. We analyzed miRNAs with a read count of 5 or more in two independent experiments. (C) RT-qPCR showing the relative levels of indicated miRNAs from non-transfected (control) or transfected (EPN-01 + Flag-MID-PIWI-Vpr) HEK293T cells. PCR reactions for each sample were carried out in triplicate. Data shown are 2^−ΔCT value expressed relative to the mean determined for the control, which was normalized to 1; results are representative of three independent experiments. Error bars represent the standard deviation. **p < 0.01, ***p < 0.001 according to Student’s t-test. Student’s t-test was performed using the Ct values of three independent experiments.