Figure 3

Specificity of hybridization between capture probes and miRNA targets in complex RNA populations. miR-199a (top panel) and miR27a (bottom panel) were captured from labeled total RNA populations prepared from 7 different RMS cell lines (RD, SMS-CTR, RH18, RH36, CCA, RH4, RH30) and a pool these RNA preparations mixed in equal amounts (POOL) with capture probes with exactly matching sequences (light grey boxes) or with one (dark grey boxes) or two mismatched bases (black boxes). The hybridization signal in all complex RNA mixtures between both capture probes and the corresponding miRNA target in comparison to the reference pooled sample shows a statistically significant decrease (p-value < 0.05) when one or two mismatches are introduced into the capture sequence. Mean value of expression and 95% confidence intervals are associated to each fluorescence value (normalized MFI represents the average of the ratio between the value of fluorescence of miRNAs and the spike).