clones6s.e.m. Construct 1; sequence 1029, Construct 2; sequence 2194. Construct 3; 780, Construct 4; lamin AC. C) Graph showing knockdown effect of shRNA construct 1 (1029) on PINK1 gene expression in individual NSC clones normalized to expression levels with vector only. Values are mean RQ; plus and minus error bars represent maximum and minimum possible RQ value. D) Western blot showing endogenous PINK1 expression in primary cortical neurons in either wild type (WT), Heterozygote (Ht) or PINK1 knockout (KO) mouse primary cortical neurons. Anti- PINK1 (Novus) was used to detect PINK expression; b-actin levels are shown as loading control. Found at: doi:10.1371/journal.pone.0002455.s003 (0.89 MB TIF) Figure S2 A) Histogram showing no significant increase in basal levels of apoptosis in PINK1 kd human NSCs compared to control cells after 24 h in culture. Levels of AnnexinV/PI staining were quantified using FACS. 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(2005) Wild-type PINK1 prevents basal and induced neuronal apoptosis, a protective effect abrogated by Parkinson disease-related mutations. J Biol Chem 280: 34025–34032. PLoS ONE | www.plosone.org 15 June 2008 | Volume 3 | Issue 6 | e2455