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. 2010 Jul 16;107(31):13794–13799. doi: 10.1073/pnas.1008684107

Fig. 3.

Fig. 3.

NLRC5 binds and transactivates MHC class I gene promoters. (A) NLRC5-mediated transactivation of MHC class I and functionally related genes. HEK293T cells were transiently transfected with either expression vectors for GFP, GFP-NLRC5, or GFP-CIITA, along with luciferase reporter constructs of the indicated gene promoters. Cell lysates were analyzed 48 h posttransfection by dual-luciferase assay. Data are a representative of three independent experiments performed in duplicates, and error bars represent ± SD. (B) Schematic representation of the W/SXY module found in the promoters of MHC class I and class II genes. The position of the primers used in the ChIP assay is indicated with arrows (P1 and P2). (C) NLRC5 occupancy, in terms of fold enrichment, at the HLA-A, -B, or -DRA promoters as determined by ChIP. Jurkat T cells stably expressing the indicated GFP fusion proteins were analyzed by ChIP assay using an anti-GFP antibody for immunoprecipitation and the indicated qPCR primers (B). GFP, empty vector; WT, wild-type NLRC5; A, Walker A mutant; B, Walker B mutant; AB, Walker AB mutant. Error bars indicate SEM (± SEM) from three independent experiments.