Figure 4: Deficiency of 4-1BB reduces inflammatory responses in skeletal muscle of HFD-fed mice. C57BL/6 wild-type (WT) and 4-1BB-deficient mice were fed a RD or HFD for 9 weeks. (a) Representative bands of 4-1BB mRNA in skeletal muscle were determined by semiquantitative RT-PCR. ((b) and (c)) TNF , IL-6, and MCP-1 mRNAs and proteins were measured by qRT-PCR and ELISA, respectively. (d) Expression of CD68 and -tubulin was determined by Western blotting using the indicated antibodies. (e) Immunohistochemical staining for macrophage-specific CD68 in paraffin-embedded sections of the quadriceps muscle. Arrows indicate CD68-stained cells. Magnification 200; Scale bars 50  m. Data are means ± SEM for . * and ** compared with WT mice. n.s., not significant.