Fig. 9. Transactivation of bone-tissue-specific marker, osteocalcin, is dependent on association of Runx2 with nuclear matrix. HeLa cells grown on six- well plates were transfected with 0.1 µg of the mentioned expression constructs along with 1 µg p208 OC CAT and 0.05 µg RSV-Luc as internal control for transfection efficiency. Cells were harvested in 300 µl of 1x Reporter Lysis Buffer (Promega) and the lysates were processed for CAT and luciferase assays as described. The CAT values were normalized with respective luciferase values and were plotted as fold activity relative to empty vector control. The graph is representative of three independent experiments each with n=6 (±s.e.m.). An example of a typical CAT assay is shown.