Fig. 7: Mechanical loading of calvarial suture cells.

A Schematic of the mechanical loading experiment. Calvarial suture cells from male 8-week-old C57BL/6J mice were seeded into gelatin-coated chambers and loaded with 5% stain at 0.5 Hz, 2 h per d for 3 d using the Cytostretcher system. Control cells were seeded in identical chambers but not subjected to biaxial strain. Gene expression was examined by qPCR 1 h after stretching or osteoblast differentiation for up to 21 d. B Mkx expression after mechanical loading by qPCR. C Expression of Mkx target genes, Sox5 and Sox6 by qPCR. D Representative images of ALP staining at 7 d (left) and statistical analysis of the staining intensity (right). E Representative images of alizarin red staining at 21 d (left) and quantitative analysis of staining (right). F Osteoblast-related genes by qPCR. G Expression of tendon-related genes by qPCR. H Expression of extracellular matrix (ECM) genes by qPCR. B, C, and F–H, data shown as mean ± 1 SD, with dots representing individual data points. D and E, individual dots in scatterplots represent values from single measurements, whereas mean and one SD are indicated by crosshairs and whiskers. All experiments were performed in triplicate replicates, with results from a single replicate shown. *p < 0.05; **p < 0.01. A two-tailed Student t-test was used for all comparisons.