Differential expression of miRNAs in MV-persistently infected UKF-NB.

<p>(A) Expression levels of selected miRNAs between uninfected and MV persistently infected UKF-NB cells. The relative expression (fold change) of each miRNA was determined by qPCR and expressed as the fold change relative to the control U6 gene. Expression in the UKF-NB cell line was used as the reference between the infected and the uninfected cells. Three independent cDNA libraries of each cell line were tested three times, each time point in triplicate. Averages with SEs are indicated for three independent experiments. (B) To determine the absolute transcript copy number per cell of miRNA-124 in UKF-NB-MV and uninfected UKF-NB cells (2X10⁵/well), we used miRNA-124 qSTAR primer pairs and a commercial miRNA-124 copy number standard kit. Averages with SEs are indicated for three independent experiments each in triplicate. T—test was performed and significance of at least p<0.05 is shown (*). (C) Relative expression of miRNA-124 in different clones of UKF-NB-MV cells. cDNA libraries of three independent UKF-NB-MV clones (I, II, III) were prepared and miRNA-124 relative expression was determined by qPCR and normalized to U6. Expression in the uninfected UKF-NB cell line was used as the reference. Triplicate samples of three independent cDNA libraries of each clone were tested. Clonal variations were observed, averages with SEs are indicated. (D) Analysis of qPCR of MV-P RNA in the different clones, normalized to U6. Expression in the uninfected UKF-NB cell line was used as the reference. Triplicate samples of three independent cDNA libraries of each clone were tested. Differences between all the clones were observed. UKF-NB-MV (fourth column) represents the parental un-cloned cells. Averages with SEs are indicated.