Figure 6.

Rmrp RNA knockdown deregulates ATDC5 chondrogenic differentiation. ATDC5 cells were transfected with 100 nM siRNA duplex targeting Rmrp RNA expression on day -1, day 2, and day 5 during chondrogenic differentiation. A scrambled siRNA was used as control condition. RNA was isolated from these cultures at day 7 and 10 in differentiation and at t = 0 as a reference. Expression of Rmrp (A), Clb2 (B), Viperin (C), ITS1 rRNA processing intermediate (D), Sox9 (E), Col2a1 (F), Runx2 (G), Col10a1 (H), Alpl (I), Bapx1 (J), PthrP (K) and Rmrp, 18S, 5.8S and 28S (L) was determined by means of RT-qPCR. Gene expression is depicted as fold induction relative to t = 0 (A–K). In Fig. 6L day 7 scrambled controls are set to 1. Data was normalized to β-actin and represents the average value of 3 (Fig. 6A–K) or 4 (Fig. 6L) biological replicates plus standard deviation. For statistical evaluation an independent samples t-test was performed relative to scrambled control for each consecutive time point using GraphPad Prism 5. p-values are indicated. Presented graphs are representative examples of three independent experiments.