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. 2011 Aug;179(2):555–563. doi: 10.1016/j.ajpath.2011.04.042

Figure 2.

Figure 2

Validation of SVV as a transcriptional target for HA–CD44. siRNA oligonucleotides specific for CD44 were transfected into MCF7F-B5 (A) and MDA-MB-231 (B) cells and CD44 (black bars) and SVV (gray bars) protein expression were determined by Western blot analysis using antibodies specific for each protein. C: CD44 induction of SVV expression is HA-dependent in MCF7F-B5 cells. Cells were cultured for 24 hours in media without doxicycline to induce the expression of CD44, after which the cells were stimulated with HA (100 μg/mL). At the indicated time points total cell extracts were performed and the expression levels of SVV were determined by Western blot analysis. In all graphs the error bars represent the SD from the mean of at least three independent determinations and the expression levels of CD44 or SVV were normalized for the β-actin loading control. Expression of either CD44 or SVV in the control sample was given a relative expression level of 1 and all other expression levels are described relative to this negative control.