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. 2013 Jun 5;288(30):22096–22110. doi: 10.1074/jbc.M113.481515

FIGURE 5.

FIGURE 5.

Cyclin D1, Runx2, and osterix mRNA levels significantly increase during osteoblast differentiation and are impacted by EB1 knockdown. A, C, and E, total RNA was collected and analyzed for cyclin D1 expression (A), Runx2 (C), or osterix (E) by quantitative RT-PCR analysis. Cyclin D1, Runx2, and osterix mRNA levels increased in MC3T3-E1 cells differentiated with AA over a 5-day time period. Each sample was done in triplicate, and data were normalized to GAPDH and are plotted as the mean ± S.E. (error bars). (*, p < 0.05, Student's t test). B, D, and F, EB1 siRNA affects cyclin D1, Runx2, and osterix mRNA expression levels. Cells were treated with AA for 5 days and transfected with EB1 siRNA or scrambled (scr) siRNA for the final 72 h. Total RNA was collected and analyzed for cyclin D1 expression (B), Runx2 (D), or osterix (F) by quantitative RT-PCR analysis. EB1 siRNA treatment significantly reduced the transcript levels of all three genes in AA-treated cells, compared with scrambled siRNA-treated cells (*, p < 0.05, Student's t test). Each sample was done in triplicate, and data are plotted as the mean ± S.E.