Myeloid-specific deletion of TAK1 prevents RANKL and LPS signaling, but not M-CSF and GM-CSF signaling.

<p>(<b>A</b>) The breeding scheme used to generate mice with TAK1 deleted in the myeloid lineage. (<b>B</b>) Western blot analysis revealed TAK1 deletion in CD11b⁺ cells. Bone marrow cells from the indicated mice were harvested, and CD11b⁺ cells were sorted by magnetic beads. Cell lysates were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. (<b>C–D</b>) Primary BMMs from TAK1^F/+ and TAK1^ΔM mice were treated with RANKL (200 ng/ml) (<b>C</b>) or LPS (200 ng/ml) (<b>D</b>) for the indicated times. Cell lysates were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. In addition, cell lysates were immunoprecipitated with anti-JNK1, and an <i>in vitro</i> kinase assay (KA) was performed. (<b>E</b>) Primary BMMs from TAK1^F/+ and TAK1^ΔM mice were treated with M-CSF (10 ng/ml) for the indicated times. Cell lysates were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. (<b>F</b>) BM cells from TAK1^F/+ and TAK1^ΔM mice were isolated, and CD11b⁺ cells were sorted by magnetic beads and treated with GM-CSF (20 ng/ml) for the indicated times. Cell lysates were subjected to SDS-PAGE and immunoblotted with the indicated antibodies. (<b>G</b>) Cell lysates from primary BMMs from TAK1^F/+ and TAK1^ΔM mice were subjected to SDS-PAGE and immunoblotted with the indicated antibodies.