Figure 2.

Knockdown of PAFAH1B2 in human cells using different RNAi approaches reduces Aβ. A, HEK293–APPsw cells transfected with non-overlapping control or PAFAH1B2-targeting siRNA pools from two sources. Efficient knockdowns of PAFAH1B2 were achieved for both pools, and this corresponded to strong reductions in Aβ secretion, by 60% for pool 1 and 40% for pool 2. Total sAPP was, on average, not significantly affected across siRNA pools. Mature (m.) and immature (im.) full-length APP and mature and immature NCT and actin were unaffected. B, SH-SY5Y cells transfected with the same non-overlapping control or PAFAH1B2-targeting siRNA pools from two sources. Efficient knockdowns of PAFAH1B2 were achieved for both pools, although not quite as good as in the HEK293 cells. This corresponded to strong reductions in endogenous Aβ secretion, by 40 and 30%, respectively. Total sAPP, full-length APP, and mature and immature NCT and actin were unaffected. C, SH-SY5Y cells transduced with non-overlapping control or PAFAH1B2-targeting shRNAs. Highly efficient knockdowns of PAFAH1B2 were achieved for both shRNAs, and this corresponded to potent reductions in endogenous Aβ secretion, by 40 and 70% respectively. Total sAPP, full-length APP, and mature and immature NCT and actin were unaffected. Con, Control; MW, molecular weight. Bar graphs show mean and SEM of six individual experiments (**p < 0.01, ***p < 0.001, unpaired Student's t test).