JCS03151 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Syntaxin 1A has a specific effect on EAAC1 surface expression. (A) The specific siRNA of syntaxin 1A suppresses the endogenous syntaxin 1A expression in the C6 glioma cells. Actin served as an internal control for protein loading. (B,C) Representative immunoblots show that syntaxin 1A has no effect on the surface expression of GLT1 (B) or EAAT4 (C) in co-transfected HEK293 cells. Actin served as an internal control for protein loading. (D) Representative immunoblot, showing that the surface expression of EAAC1 is not regulated by co-transfection of syntaxin 4, SNAP-25 or VAMP-2 in HEK293 cells. Actin served as an internal control for protein loading.

• Supplemental Figure 2 -

  Fig. S2. The effect of specific syntaxin 1A small interference RNA (siRNA) on EAAC1 surface expression and internalization. (A) In contrast to NS siRNA, transfection of Syn1A siRNA leads to an increase in EAAC1 expression in both total lysate and cell surface fraction. **P<0.01 compared with cells transfected with NS siRNA (n=3). (B) Representative immunoblot and quantitative analysis of internalized EAAC1, showing that the internalization of EAAC1 is impaired after the suppression of endogenous syntaxin 1A in C6 glioma cells, despite of Syn1A siRNA increasing the overall EAAC1 pools available for internalization. The band densities for the internalized EAAC1 are plotted as a percentage of the total surface EAAC1 available for internalization within each transfection group. *P<0.05, **P<0.01 versus the cells transfected with NS siRNA at the same time point (n=3).