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. 2017 Oct 31;292(51):20883–20896. doi: 10.1074/jbc.M117.809517

Figure 7.

Figure 7.

Icariin-induced activation of cAMP/PKA/CREB signaling and osteogenesis in rat calvarial osteoblasts require the existence of primary cilia. A, the mRNA and the protein expression levels of IFT88 in cells transfected by negative control siRNA (NC group) or IFT88 siRNA (siRNA group). B, the percentage of cells with primary cilia after the transfection. C, primary cilia of the transfected osteoblasts. The primary cilium was stained with acetylated α-tubulin (green), and DNA stained with DAPI (blue). Bar = 20 μm. D, the intracellular cAMP contents in the transfected osteoblasts after icariin treatment for 120 min. E, protein abundance of AC2, AC9, and sAC, p-PKA, p-CREB, total PKA, and CREB in the transfected osteoblasts after icariin treatment. F, the nuclear translocation of p-PKA in the transfected osteoblasts after exposure to icariin. G, ALP activity in the transfected osteoblasts after 3 and 6 days of icariin supplement. H and I, the relative mRNA expression levels of ALP, COL1α2, and RUNX2 (H) and protein expression levels of COL1 and RUNX2 (I) in transfected osteoblasts after 3 and 6 days of icariin supplement. Each experiment was conducted at least three times independently. Data are represented as mean ± S.D. (n = 3); *, p < 0.05; **, p < 0.01; ***, p < 0.001 when compared with NC or NC + icariin group.