Research Paper Volume 13, Issue 3 pp 4199—4214

Figure 1. Endogenous expression of HOTAIR in different tissues of mice and in MSCs undergoing osteogenic differentiation. (A) Bones exhibit higher expression of HOTAIR than most other tissues. Total RNA from different tissues of 4-week-old male CD1 mice (N=5) was extracted and subjected to reverse transcription followed by TqPCR analysis. Reactions were done in triplicate. “##” p > 0.05, bones vs spleen, “*” p < 0.05, bones vs other tissues/organs. (B) Expression of HOTAIR in femurs was peaked at 4-week, a fast growing period for mice skeleton. Total RNA from different ages of CD1 mice femurs (N=5) was extracted and subjected to reverse transcription followed by TqPCR analysis. Reactions were done in triplicate. “*” p < 0.05, 4-week vs 2-week/3-month. (C) Endogenous expression of HOTAIR in iMADs is up-regulated during osteogenic differentiation and quickly dropped in the late stage, and BMP9 could potentially bring forward the peak of HOTAIR expression. Subconfluent iMAD cells were infected with Ad-GFP or Ad-BMP9. Total RNA was isolated at 48 h 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 GFP group vs iMAD BMP9 group.