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. 2007 Mar 13;96(7):1062–1071. doi: 10.1038/sj.bjc.6603672

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Copyright © 2007 Cancer Research UK

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Apoptosis in SH-SY5Y cells (A) and A375 cells (B) after gene knockdown with control, ERdj5-2 or ERp57-3 siRNA. Apoptosis in response to control vehicle, fenretinide (3 or 10 μM), velcade (5 or 30 nM), vincristine (10 nM) or temozolomide (1 mM) (SH-SY5Y or A375 cells, respectively), or thapsigargin (1.5 μM for SH-SY5Y or 7.5 μM for A375 cells) was measured quantitatively by flow cytometry of PI-stained cells; the detection of cleaved caspase-3 on Western blots (a technique that is less sensitive than flow cytometry), relative to β-actin as a loading control, was used as additional verification of apoptosis (upper panels in A and B). Confirmation of ERp57 and ERdj5 protein knockdown was assessed by Western blotting. Bar heights indicate the mean with error bars showing the upper 95% confidence limits. Statistical comparisons were hypothesis tests (Bonferroni-corrected) from within a two-way ANOVA in which the effect of siRNA knockdown of ERdj5 or ERp57 was compared with the scrambled control for that drug treatment; ^*P<0.05; ^***P<0.0001.