FIGURE 10.

siRNA-mediated knockdown of BiP reduces the steady state levels of torsinA and torsinAΔE. HeLa cells were treated with siRNA against BiP for a total of 48 h and were transfected with torsinA (T) and torsinAΔE (ΔE) expression vectors 24 h before harvest. A, representative Western blot; B, graph showing the average ± S.E. of the normalized levels of BiP, torsinA, and torsinAΔE after treatment with a control siRNA or siRNA directed against BiP (077). Experiments were performed at least twice with three replicates per experiment. The graphed values were normalized as follows. The levels of BiP, torsinA, and torsinAΔE for each replicate in each experiment were normalized first to the corresponding actin levels. Then the actin-normalized value from one siRNA control replicate was used to normalize the values of all other replicates. This procedure was performed for each torsinA and torsinAΔE experiment independently. Actin was used as loading control. Grp94 levels were also monitored by Western blot.