spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.



Fig. 9. Analysis of glycosylation of tagged Syt I and II. (A) RBL cells transiently transfected with T7-Syt I, HA-Syt II or the chimeras HA-Syt I/Syt II and HA-Syt II(1-183aa)/Syt I cDNA were grown for 2 hours at 37°C and subsequently left untreated or incubated for 24 hours in the presence of tunicamycin (10 µg/ml). Cell lysates were resolved by SDS-PAGE and probed with either anti-T7 or anti-HA antibodies, as indicated. (B) Cell lysates derived from RBL cells transfected with T7-Syt I, HA-Syt II, HA-Syt I/Syt II and HA-Syt II(1-183aa)/Syt I cDNA were treated with N-glycosidase F or buffer as described in Materials and Methods. Lysates were subjected to SDS-PAGE and probed with anti-T7 or anti-HA antibodies. (C) Cell lysates (80 µg) derived from RBL cells stably transfected with Syt II-GFP or Flag-Syt II-GFP were treated with N-glycosidase F, resolved on SDS-PAGE and probed with anti-GFP antibodies. The asterisks in panels A-C indicate proteins that were reduced by tunicamycin or N-glycosidase F treatment. (D) RBL cells transfected with Syt II(T34/A)-GFP or Flag-Syt II(T34/A)-GFP were subjected to tunicamycin or (E) N-glycosidase F treatment as described above. (F) Lysates (80 µg) derived from RBL cells transiently transfected with Flag-Syt II-GFP (WT)(1, 5) or Flag-Syt II(T34/A)-GFP ({Delta}N)(2), or Syt II-GFP (WT)(3) or Syt II(T34/A)-GFP ({Delta}N)(4) or Flag-Syt II(T17/18/19/A)-GFP ({Delta}O) (6) were subjected to SDS-PAGE and probed with anti-GFP antibodies. Lane 3 is a longer ECL exposure time of the lane marked 3*.





Right arrow Return to article