Figure 1.

(A) CD4+ T-cells were isolated from PBMC of three donors (1–3) with positive (+) or negative “untouched” (−) bead-based selection. OpenArray profiling data for miRNAs amplifying before relative threshold cycle (Crt) of 30 were quantile normalized and clustered using Pearson correlation and average linkage. Correlation scale on right indicates relatedness of samples. (B) No individual miRNA was significantly differentially regulated following application of multiple comparison correction to paired t-tests (OpenArray). Shown are two miRNAs with uncorrected and nominally “significant” p values (uncorrected p<0.05 in data normalized by at least one technique of quantile normalization, geometric mean of all Crt<30 (“GM All”), geometric mean of U6, RNU44 and RNU48 (“GM 3”) or miR-16. (C) Individual qPCR assays confirmed a lack of significant differences between positively and negatively selected CD4+ T-cells for candidate miRNAs (from OpenArray) or for miRNAs previously reported to be differentially expressed following cell activation. (D) As a positive control, significant differences were confirmed for several miRNAs following activation of CD4+ T-cells anti-CD3/anti-CD28 antibodies. For C and D, statistical significance was assessed for positively and negatively selected cells or stimulated versus unstimulated cells by Students t-test and indicated as follows: p<0.05 (*), p<0.01 (**), p<0.001 (***). All features marked with multiple asterisks had statistically significant differences following Bonferroni correction for multiple tests. Raw data and quantile normalized processed data have been uploaded to the Gene Expression Omnibus.