To determine osteoblastic differentiation of MC3T3-E1 cells, the ALP activity of the cells was assayed with the ALP kit mentioned above, the protein level of Collagen type I (Col-I) was detected via Western blot, and the mRNA levels of osteocalcin (OCN) and Runx2 were assayed via RT-qPCR, The methods of Western blot and RT-qPCR were described above, the primers of OCN were as follows, forward:
AGTCTGACAAAGCCTTCA, reverse:AAGCAGGGTTAAGCTCACA; the primers of Runx2 were as follows, forward: AGTAGCCAGGTTCAACGAT, reverse: GGAGGATTTGTGAAGACTGTT. In addition, the Bone morphogenetic protein-2 (BMP-2) protein level in the culture supernatant of the cells was detected via ELISA also mentioned above.
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