Figure 2. β-Glucocerebrosidase knockdown does not raise Gb3.
(A) Knockdown of the GBA gene and suppression of β-glucocerebrosidase, another lysosomal hydrolase, in cultured EA.hy926 cells by three duplexes of anti-human GBA-dsiRNA (A, B and C) at the indicated exposure times as confirmed by immune bot analysis. (B) Lipids analysis of the GlcCer levels in control-dsiRNA and GBA-dsiRNA transfected EA.hy926 cells at days 3 and 4 of following a single transfection (ST) and at day 6 following a double transfection (DT). Abbreviations include GalCer, galactosylceramide; Std., standards. (C) Determination of GlcCer accumulation in control- and GBA-dsiRNA transfected cells by densitometric scanning (n = 3). (D) Levels of Gb3 in GBA-siRNA transfected cells assessed in parallel by high performance thin layer chromatography, and (E) as determined by scanning densitometry (n = 3). The error bars represent ± SE.



