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Blood, Vol. 109, Issue 8, 3385-3392, April 15, 2007
Synaptotagmin (Syt) IX is an essential determinant for protein sorting to secretory granules in mast cells
Blood Haberman et al.
109: 3385
Supplemental materials for: Haberman et al, Vol 109, Issue 8, 3385-3392
Files in this Data Supplement:
- Figure S1. Characterization of Syt IX-overexpressing or knocked-down cells (PDF, 98.4 KB) -
(A) Control, RBL-Syt IX−, and RBL-Syt IX+ cells were labeled with polyclonal anti–mannosidase II antibodies, followed by Cy3-conjugated donkey antirabbit antibodies. Bars represent 10 µm. (B) Cell lysates (80 µg) derived from control, RBL-Syt IX+, or RBL-Syt IX− cells were separated by SDS-PAGE and analyzed by Western blot using the indicated antibodies.
- Figure S2. Expression of HA-TGN38 in RBL clones (PDF, 66.3 KB) -
Cell extracts (80 µg) derived from control, RBL-Syt IX−, or RBL-Syt IX+ cells transiently transfected with HA-TGN38 cDNA were resolved by SDS-PAGE and subjected to immunoblotting with monoclonal anti TGN38 antibodies, anti-HA antibodies, and antitubulin.
- Figure S3. Biosynthetic trafficking of transiently transfected HA-TGN38 in control, Syt IX knocked-down, or Syt IX-overexpressing RBL cells (PDF, 210 KB) -
Control, RBL-Syt IX−, and RBL-Syt IX+ cells were transiently transfected with HA-TGN38 cDNA and grown on glass coverslips for 1 hour at 37°C. Cells were then shifted to 20°C, incubated for 2 hours to accumulate proteins at the Golgi complex, and then delivered to 37°C for the indicated time periods. Cells were either left untreated (A) or treated with brefeldin A (BFA; 5 µg/mL) for the last 30 minutes of the indicated incubation time (B). Cells were subsequently labeled with monoclonal anti-HA antibodies followed by Cy3-conjugated donkey anti–mouse IgG. Bars represent 3 µm.
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