Ocn mm03413826

Quantitative gene expression analysis was carried out as we previously described [48 (link)]. Briefly, 1×10⁵ C2C12 cells were seeded into GF/MSN, GF/MSN/CTS-DFO, GF/MSN-BMP2 and GF/MSN-BMP2/CTS-DFO scaffolds. After 7 days of incubation, total RNA was extracted using the GeneJET RNA Purification Kit (Thermo Scientific, Waltham, MA) by following the manufacturer’s instruction. An equivalent amount of RNA was processed to generate cDNA by using the High Capacity cDNA Reverse Transcript kit purchased from Applied Biosystems (Forster City, CA). Quantitative PCR was performed with Taqman gene expression assays (Applied Biosystems, Forster City, CA) using the Applied Biosystems 7500 Fast Real- Time PCR System (Applied Biosystems, Carlsbad, CA). TaqMan^® Gene Expression Assays of GAPDH (Mm99999915), RUNX2 (Mm00501584), BSP (Mm00436767) and OCN (Mm03413826) were purchased from Applied Biosystems (Forster City, CA).

Quantitative gene expression analysis was carried out as we previously described (Yao et al., 2016 (link)) with some minor modifications. Briefly, total RNA was extracted using the GeneJET^™ RNA Purification Kit (Thermo Scientific^™, Waltham, MA) by following the manufacturer’s instruction. RNA concentration was measured by UV–Vis spectroscopy (DU 730, Beckman coulter) at 260 nm and an equivalent amount of RNA was processed to generate cDNA by using the High Capacity cDNA Reverse Transcript kit purchased from Applied Biosystems (Forster City, CA). Quantitative PCR was performed with Taqman gene expression assays (Applied Biosystems, Forster City, CA) using the Applied Biosystems 7500 Fast Real-Time PCR System (Applied Biosystems, Carlsbad, CA). Triplicates were performed for each sample, and results were normalized to GAPDH. TaqMan® Gene Expression Assays of GAPDH (Mm99999915), RUNX2 (MmCG122221), BSP (Mm00436767), and OCN (Mm03413826) were purchased from Applied Biosystems (Forster City, CA).