FIG 1.

Generation of myeloid cell-specific NFI-A knockout model. (A) Schematic representation of gene targeting method used in the generation of the myeloid cell-specific NFI-A knockout mice. (B) The locations and sequences of the PCR primers used to confirm the gene targeting and sizes of the expected PCR products are shown. WT, wild type. (C) Nfia genotyping. The DNA was isolated from tail samples or Gr1⁺ CD11b⁺ cells and analyzed by PCR using the primers shown in panel B. PCR of the floxed (wild-type) allele produced a 405-bp product from tail DNA samples or Gr1⁺ CD11b⁺ cells, whereas PCR of the deleted allele produced a 577-bp product from Gr1⁺ CD11b⁺ cell DNA from the conditional knockout only, confirming successful deletion of Nfia in the myeloid lineage. f, flox. (D) NFI-A protein levels. Gr1⁺ CD11b⁺ cells were isolated from the bone marrow of the control (Nfia^flox/flox;Lyz2^+/+) and NFI-A-deficient (Nfia^flox/flox;Lyz2^cre/+) mice undergoing sepsis, and levels of the NFI-A protein were measured by Western blotting. The results are representative of two immunoblots from two independent experiments. cKO, knockout; Flox, floxed allele; Del, deleted allele.