FIGURE 10.

BMP2 promotes the invasive differentiation of mouse TS cells. (A) TS cells were treated with Ctrl or 100 ng/mL BMP2 every 24 h for up to 7 days, and three cell marker genes, including Eomes (TS cell marker), Tpbpa (SpT marker) and Ctsq (TGC marker), were examined using RT-qPCR. (B) TS cells were treated with Ctrl or 100 ng/mL BMP2 for 4 days, and cell images were detected with microscopic observation. Undifferentiated TS cells showed a colony with characteristic tightly packed cells (yellow arrowheads), while differentiated TS cells showed a typical TGC morphology (blue arrowheads). These differentiated TS cells showed a flattened appearance and increased cell, nuclei and perinuclear granule sizes (blue arrowheads). The scale bars represent 200 μm. (C) TS cells were treated with Ctrl or 100 ng/mL BMP2 every 24 h for a total of 72 h, and cell invasion was examined using a Matrigel-coated transwell invasion assay. Cell viability was determined using a CCK-8 assay. (D) TS cells were treated for 12 or 24 h with Ctrl or 100 ng/mL BMP2, and the mRNA (12 h) and protein (24 h) levels of MMP2 and SNAIL were examined using RT-qPCR and western blot analyses, respectively. TS cells were treated for 24 h with Ctrl or 100 ng/mL BMP2, and the MMP2 activity was examined using ELISA. GAPDH and α-tubulin were used to normalize the RT-qPCR and western blot results, respectively. The results are expressed as the mean ± SEM of three independent experiments (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.001). Differences between groups were determined by Student’s t-test or one-way analysis of variance.