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. Author manuscript; available in PMC: 2009 Jan 8.
Published in final edited form as: Biochemistry. 2007 Dec 11;47(1):143–156. doi: 10.1021/bi701559f

Fig.7. Potentiation of the 4-HNE-cytotoxicity to CRL2571 cells upon silencing of Daxx with siRNA.

Fig.7

(A) Silencing of Daxx: Cells were transfected with Daxx siRNA duplex (50pmol) as per manufacturer's instructions (SantaCruz Biotech) while controls were treated with scrambled siRNA duplex in a similar manner. The treated cells were harvested 48h post transfection and silencing of Daxx expression was examined by Western blot analysis of the cell extracts using anti-Daxx antibodies. Daxx siRNA (middle lane) caused almost complete silencing of Daxx expression. After ascertaining silencing of Daxx expression, Daxx-depleted cells (2×105) were then grown in six well plates and treated with 10μM 4-HNE for 2h. (B) Phase contrast micrographs showing the effect of 4-HNE (100×) and (C) bar graph showing the percent cell survival determined by Trypan blue dye exclusion, of Daxx depleted and scrambled siRNA transfected cells in control and 4-HNE treated cells. Values are mean ±SD of two separate experiments done in triplicate (* significant differences observed in control and treated cells p<0.01)