Figure 2. Construction and verification of siRNA targeting mouse HOTAIR in iMADs. (A) Schematic configuration of the three tandem targeting siRNAs (simHOTAIR1, 2, and 3) and their targeting sequences and locations on mouse HOTAIR gene. (B) Efficient silencing of endogenous mHOTAIR expression in iMADs. Total RNA was isolated from subconfluent iMADs that were stably transduced with the pSOK vector expressing three simHOTAIR sites (iMAD-KD) or scrambled controls (iMAD-Ctrl) at Day 1 and 3 post transduction, and subjected to TqPCR analysis using gene-specific primers for mouse HOTAIR. Gapdh was used as a reference gene. Reactions were done in triplicate. “*” p < 0.05, iMAD-Ctrl group vs iMAD-KD group.