Real-time RT-PCR analyses of hypertrophic markers in transduced lines (day 28) cultured under nonmineralizing conditions. RNA levels were normalized to expression of β-actin. (a) Expression of col10a1 in transduced lines. Expression is higher than empty vector in cells overexpressing wild-type (WT) ank (*P ≤ 0.05); however, in cells expressing mutant ank, expression levels of col10a1 are equivalent to empty vector. (b) Expression of osteopontin in transduced lines. Expression of all transduced cells is lower than empty vector, but not significantly different among cell lines that express mutant ank versus cell line overexpressing WT ank. (c) Expression of runx2 (cbfa1) in transduced lines. Expression is equivalent for all transduced lines and not significantly different from the expression of runx2 in cells transduced with empty vector only. n = 6. At least three independent clones for each cell line were evaluated; results presented are from single clones and are representative of other clones for each cell line.