|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 7. Selective decrease of Syt V and Syt IX expression by RNA interference. (A) INS-1E cells were transiently transfected with myc-tagged Syt V (a) or myc-tagged Syt IX (b). Silencing of the genes was determined by cotransfecting the cells either with an empty pSilencer vector (control) or with a pSilencer vector containing a sequence that directs the synthesis of Syt V-specific siRNA or Syt IX-specific siRNA (RNAi). Nontransfected cells are shown by (–). After three days the cells were homogenized and equal amounts of protein were analysed by western blotting using antibodies directed against Syt V or Syt IX. The same membrane was also blotted with an anti-myc antibody. The arrow indicates Syt IX unrelated proteins. (B) INS-1E cells were transiently cotransfected with GFP and with an empty vector (control) or with a vector containing either the Syt V or the Syt IX silencer sequences (RNAi). Three days later, GFP-expressing cells were enriched by FACS separation. The cells were then homogenized and the same amount of protein was separated by SDS-PAGE and immunoblotted with anti-Syt V (a) or anti-Syt IX (c) antibodies. The membranes (a) and (c) were then blotted respectively with anti-Syt IX (b) and anti-Syt V (d) antibodies.
|
