Prevention of type 1 diabetes by gene therapy
J. Clin. Invest. Chaorui Tian, et al. 114:969 doi:10.1172/JCI22103 [
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Figure 4Subcellular localization of retrovirally encoded I-Aβ chains. (
A) Retrovirally encoded I-Aβ chains are expressed on the surface of MHC class II_positive cells. PBMCs were harvested from NOD mice reconstituted with either MMP-GFP_transduced (top panels) or MMP-IAβ-d-GFP_transduced bone marrow (bottom panels), fixed, and stained with antibodies specific for GFP (left panels, green) and I-A
d (middle panels, red). The anti_I-A
d antibody used cross-reacts with I-A
g7. Right panels: Overlay images showing colocalization of retrovirally encoded I-A
d_GFP fusion proteins with endogenous I-A
g7 on the surface in yellow. (
B) Retrovirally encoded I-Aβ chains are expressed in the cytoplasm of surface-MHC class II negative cells. PBMCs from NOD mice reconstituted with MMP-IAβ-d-GFP_transduced bone marrow were fixed and stained with antibodies specific for GFP (left panel, green) and anti-CD3 (middle panel, red). An overlay of the 2 images is shown in the right panel, demonstrating that I-Aβ
d_GFP does not colocalize with CD3 on the cell surface of MHC class II_negative cells. Shown are representative results of 3 independent experiments.