TUT1 knockdown with additional, independent siRNAs confirms the role of TUT1 in maintaining miRNA expression levels.

<p>(<b>A</b>) Two additional TUT1 siRNAs were used to suppress TUT1 in HCT-116 cells. Bar graphs show qRT-PCR assessement of the suppression of TUT1 compared to cells treated with negative control siRNAs. (<b>B</b>) RNAi with both siTUT1-A and siTUT1-B decreased the expression of the TUT1 protein in HCT-116 cells. (<b>C</b>) Exiqon miRNA qRT-PCR arrays were used to compare the global miRNA expression in enzyme suppressed versus negative control cells. Boxplots depict miRNA expression following TUT1 suppression with each of the two new TUT1 siRNAs. Points below the dashed line indicate decreased miRNA abundance with TUT1 suppression compared to the negative control cells. (<b>D</b>) Bar graphs compare the expression of two small nucleolar RNAs, SNORD38B and SNORD49A, in TUT1 supppressed versus control cells. TUT1 suppression did not affect small nucleolar RNA abundance levels, which confirms the specificity of the phenotype to miRNAs.