Nse: 59-UUC UCC GAA CGU GUC ACG UTT-39; antisense: 59-ACG UGA
Nse: 59-UUC UCC GAA CGU GUC ACG UTT-39; antisense: 59-ACG UGA CAC GUU CGG AGA ATT-39. Briefly, MC3T3-E1 cells were grown in a-MEM devoid of antibiotics prior to siRNA treatment. The transfection medium was replaced just after five h. Protein assays to assess knockdown were performed at 48 and 72 h just after transfection. Functional assays have been performed through maximum knockdown61,62. Synthesis and transfection of miRNA inhibitor. The miR-103 CA XII Inhibitor Biological Activity inhibitor was designed and synthesized by RiboBio Corporation. The sequence of miR-103 inhibitor is 3′-UCA UAG CCC UGU ACA AUG CUG CU-5′. 5 nucleotides or deoxynucleotides at each ends on the antisense molecules had been locked. Osteoblasts were transfected with inhibitor or unfavorable manage using Lipofectamine 2000. The medium was replaced at 6 h after transfection. The cells had been collected for protein assay or patch clamp at 48 h just after transfection35. 1. Duncan, R. L. Turner, C. H. Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tissue Int 57, 34458 (1995). two. Nishizuka, Y. Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 258, 60714 (1992). three. Riggs, B. L., Khosla, S. Melton, L. R. A unitary model for involutional osteoporosis: estrogen deficiency causes each type I and form II osteoporosis in postmenopausal ladies and contributes to bone loss in aging men. J Bone Miner Res 13, 76373 (1998). 4. Yagodovsky, V. S., Triftanidi, L. A. Gorokhova, G. P. Space flight effects on skeletal bones of rats (light and electron microscopic examination). Aviat Space Environ Med 47, 73438 (1976). 5. Morey, E. R. Baylink, D. J. Inhibition of bone formation throughout space flight. Science 201, 1138141 (1978). six. Jee, W. S., Wronski, T. J., Morey, E. R. Kimmel, D. B. Effects of spaceflight on trabecular bone in rats. Am J Physiol 244, R310 314 (1983). 7. Wronski, T. J. Morey, E. R. Impact of spaceflight on periosteal bone formation in rats. Am J Physiol 244, R305 309 (1983). eight. Zerath, E. et al. Effects of spaceflight on bone mineralization in the rhesus monkey. J Appl Physiol (1985) 81, 19400 (1996). 9. Patterson-Buckendahl, P. et al. Fragility and composition of ERK2 Activator custom synthesis expanding rat bone soon after one particular week in spaceflight. Am J Physiol 252, R240 246 (1987). ten. Doty, S. B., Morey-Holton, E. R., Durnova, G. N. Kaplansky, A. S. Morphological studies of bone and tendon. J Appl Physiol (1985) 73, 10S3S (1992). 11. Zerath, E. et al. Spaceflight inhibits bone formation independent of corticosteroid status in expanding rats. J Bone Miner Res 15, 1310320 (2000). 12. Vico, L. et al. Effects of long-term microgravity exposure on cancellous and cortical weight-bearing bones of cosmonauts. Lancet 355, 1607611 (2000). 13. Landis, W. J., Hodgens, K. J., Block, D., Toma, C. D. Gerstenfeld, L. C. Spaceflight effects on cultured embryonic chick bone cells. J Bone Miner Res 15, 1099112 (2000). 14. Pardo, S. J. et al. Simulated microgravity using the Random Positioning Machine inhibits differentiation and alters gene expression profiles of 2T3 preosteoblasts. Am J Physiol Cell Physiol 288, C12111 (2005). 15. Bergh, J. J., Shao, Y., Puente, E., Duncan, R. L. Farach-Carson, M. C. Osteoblast Ca21 permeability and voltage-sensitive Ca21 channel expression is temporally regulated by 1, 25-dihydroxyvitamin D3. Am J Physiol Cell Physiol 290, C822 831 (2006). 16. Bergh, J. J., Shao, Y., Akanbi, K. Farach-Carson, M. C. Rodent osteoblastic cells express voltage-sensitive cal.
