**3.7 SiRNA effect on HDAC 8 and C MYC**

HDAC 8 and c MYC proteins were quantitated by antibody in ELISA in cell extracts prepared from the cells transfected with negative control siRNA and siRNA-HDAC 8. The results show that siRNA-HDAC 8 transfection inhibited 40% of HDAC 8 proteins (**Figure 5B**) as well as inhibited 35% of MYC protein. Inhibition of MYC may be indirect effect of HDAC 8 inhibition. HDAC8 inhibition increases acetylation of histones and alters gene expression, thus decreasing MYC expression. Therefore, miR-665 represses the expression of c MYC both at the transcription and at the post transcription levels. siRNA-HDAC 8 and siRNA-c MYC substantiated the effects of miR-665 on neuroblastoma cells.

## **3.8 siRNA activates caspase 3**

Caspase 3 activity was measured in the cell extracts prepared from cells transfected with negative control siRNA, siRNA-HDAC8 and siRNA-c MYC. The results show that siRNA-HADC8 increased the activity of caspase 3 by 1.8-fold and siRNA –c MYC increased the activity of caspase 3 by 2.5-fold compared to negative control siRNA (**Figure 5C**). Therefore, the results of siRNA effects substantiate the effects of miR-665 on the activation of caspase 3.
