FIG 4.

Dominant negative HES1 upregulates DTX1 expression. (A) The HES1-AB reporter is activated by CSL/NOTCH and inhibited by HES1. 293T cells were cotransfected with HES1-AB reporter (firefly luciferase) and a Renilla luciferase control together with various dosages of HES1 and/or dnHES1 as shown in the figure. Relative luciferase activity was measured 48 hours after transfection. (**, p<0.005) (B) PCR and (C) quantitative PCR analysis of changes in HASH1 and DTX1 expression induced by dnHES1. Osteosarcoma cells OS 187, COL and KRIB were transduced with empty vector (V) or dnHES1 (dnH). Gel image (B) shows the PCR amplification of HASH1, DTX1 and actin; histograms (C) represent relative Q-PCR results, normalized to a value of 1 for the vector-only cells. (D) Histograms represent matrigel invasion of the same cells described in 4B and 4C. Analysis was performed as in figure 1E. (**, p<0.005. *, p<0.05). V: empty vector control. dnH: dnHES1.