Nature research paper: Autoimmunity in Down’s syndrome via cytokines, CD4 T cells and CD11c+ B cells
Overnight, 96-well Costar plates were coated at 4 °C with 100 μl per well of a 1 μg mlsolution of recombinant IFNGR2, MSTN or ATP6V1G2 protein suspended in 1× PBS. The next morning, the coating solution was removed and wells were washed three times with 100 μl of washing buffer Tween 20; PBS-T). Next, 200 μl of blocking buffer ) was added to each well at room temperature and incubated at room temperature for 1 h. Plasma samples were diluted 1:100 in blocking buffer.
. HEK293T cells were transfected with a plasmid containing the firefly luciferase gene under the control of the human ISRE promoter in the pGL4.45 backbone and a plasmid constitutively expressingluciferase for normalization . Cells were transfected in the presence of the X-tremeGENE9 transfection reagent for 24 h.
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