|
|
Blood, Vol. 113, Issue 15, 3568-3576, April 9, 2009
Anchoring of FLT3 in the endoplasmic reticulum alters signaling quality
Blood Schmidt-Arras et al.
113: 3568
Supplemental materials for: Schmidt-Arras et al
Files in this Data Supplement:
- Figure S1. Effect of the kinase inhibitor PKC412 on FLT3 dependent activation of STAT3 and STAT1 proteins (JPG, 41.5 KB)
-
32D cells expressing wildtype FLT3 R3, FLT3 A3, or FLT3 ITD were starved, and treated with 30 nM PKC412 for 4h, before analysis of cell lysates by immunoblotting.
- Figure S2. Dephosphorylation of ER-anchored FLT3 by the protein-tyrosine phosphatase PTP1B (JPG, 50.5 KB)
-
Immortalized mouse embryo fibroblasts with inactivated PTP1B gene (PTP1B −∕−), or restored with wildtype PTP1B (PTP1B WT) were transfected with expression constructs encoding the indicated untagged or ER-anchored (R3) FLT3 versions. FLT3 was immunoprecipitated, and tyrosine phosphorylation was analyzed by immunoblotting using anti-phosphotyrosine antibodies (4G10). The blots were stripped and reprobed for FLT3.
- Figure S3. Effect of signal transduction inhibitors on proliferation of 32D cells expressing different FLT3 mutants (JPG, 96 KB)
-
32D cell lines expressing the indicated FLT3 variants (D-Y, FLT3 D835Y untagged; D-Y A3, FLT3 D835Y A3; ITD, FLT3 ITD untagged) were seeded in 96-well plates (2.5 × 104 cells per well), signal transduction inhibitors were added as indicated (final DMSO concentration 0.1 %), and cell growth in the absence (left panels) or presence of IL-3 (2.5 ng/ml, right panels) was measured after 72 hours using the MTT method. Values were normalized to growth with solvent controls, which were set to 100%.
|
|
