Figure 5.

F4/80⁺ cells express macrophage markers. Perigonadal adipose tissue was collected from female B6.V Lepob/ob mice, digested, and centrifuged to yield a buoyant adipocyte-enriched cell population and a pellet of SVCs. The SVCs were separated into F4/80⁺ (black bars) and F4/80– (white bars) populations via FACS. Quantitative RT-PCR was used to measure the relative expression of macrophage markers (Emr1, Csf1r, Cd68) and an adipocyte-specific gene (Acrp30). Among the three isolated cell populations, the relative gene expression of macrophage markers was highest among the F4/80⁺ cells. *The F4/80^– SVCs did not express detectable amounts (< 0.05 of mean of all populations) of the macrophage markers. The adipocyte-enriched population (gray bars) expressed small amounts of the macrophage markers (a), consistent with residual macrophage contamination seen by immunofluorescent staining of live cells (b). In the adipocyte-enriched fraction, large autofluorescent adipocyte cell membranes (green) are not recognized by fluorescently conjugated F4/80 antibody (red), but membrane staining of small nonautofluorescent cells is seen. Control fluorescently conjugated isotype antibody did not recognize these cells (c).