The RT-PCR results showed that the expression of ASB16-AS1 transcript variant 1 was higher than the expression of AB16-AS1 transcript variant 2 (Fig. 5A). However, using the same primers, we failed to get a clone of variant 1. The variant 2 was successfully subcloned into the pCEP4 vector. After transfection of pCEP4-ASB16-AS1, the expression of ASB16-AS1 were highly enhanced (Fig. 5B). The expression of BMP2 and ALPL was significantly increased in the ASB16-AS1 over-expressed osteoblast compared with the control (Fig. 5C and D). The expression of RUNX2 was increased but the alteration was not significant (Fig. 5E).
