Fig. 6

Identification of s100a10 as a uniquely expressed mineralization gene using in vitro comparative RNA-seq. a S100A10 immunofluorescence staining in regenerating deer antler tissue. b RM cells (isolate 2) cultured with 100 ng/mL BMP-2 and 100 nM dexamethasone exhibited increased S100A10 expression relative to control. c C3H10T1/2 cells stably transfected with s100a10 and cultured with 100 ng/mL BMP-2 for 4 h exhibited increased alp gene expression relative to untransfected control and empty plasmid control. C3H10T1/2 cells stably transfected with s100a10 and cultured with 100 ng/mL BMP-2 for 12 days exhibited increased ocn and runx2 gene expression relative to their respective control. d C3H10T1/2 cells stably transfected with s100a10 and cultured with 100 ng/mL BMP-2 for 4 days exhibited increased ALP activity relative to untransfected control. e C3H10T1/2 cells stably transfected with s100a10 and cultured in the presence of 100 ng/mL BMP-2 and 100 nM dexamethasone exhibited increased Alizarin Red S staining relative to untransfected control and empty plasmid control. Scale bars as indicated. Data were from n = 3 isolates (an independent herd for antler immunofluorescence studies), n = 2–3 independent experiments with 4–10 replicates per group for osteogenic gene expression studies, n = 3 independent experiments with 9 replicates per group for s100a10 overexpression ALP studies, and n = 5 independent experiments with 15 replicates per group for s100a10 overexpression mineralization studies. Gray circles indicate observed data points. Error bars indicate SEM. Statistical significance as indicated