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. Author manuscript; available in PMC: 2010 Aug 1.
Published in final edited form as: Biochim Biophys Acta. 2009 Apr 2;1791(8):730–739. doi: 10.1016/j.bbalip.2009.03.012

Figure 5. Effectiveness and specificity of NCDase siRNA.

Figure 5

Cells were transfected with scrambled siRNA or NCDase siRNA for about 36 hours and then cells were collected, lysed and then subjected to RT-PCR, Western and NCDase activities assays as described under “Materials and Methods”. (A) Real time RT-PCR analysis of NCDase mRNA level. (B)Western blot analysis of endogenous NCDase expression in H. end. cells. Equal loading was verified by using anti-β-actin. (C) TLC analysis of NCDase activity in NCDase siRNA treated cells. (D) NCDase activity was quantified by densitometry. The values are expressed as the mean ±SD (n=3). Results are representative of three independent experiments. SCR, scrambled siRNA; NCD, NCD siRNAi; FA, fatty acid product; NBD CER, NBD C-12 ceramide substrate. Statistical significance was calculated with respect to scrambled control (*, p<0.05)