Figure 2

Characterization of adipocyte preparations isolated from bone marrow and epididymal depots. A. Light and immunofluorescence microscopy of adipocytes isolated from bone marrow and epididymal white adipose tissue. Fixed cells were incubated with either BODIPY 493/503 (1:500) or PE (phycoerythrin) conjugated anti-mouse CD11b (Intergrin a, Mac-1α) and polyclonal anti rabbit perilipin antibodies (1:200) in 1% blocking solution for 1 h. Following 1 h incubation of Alexa 555 (red) conjugated secondary antibodies at a dilution of 1/800 at room temperature, the stained cells were washed three to four times with PBS and observed using a Zeiss microscope observer A1. B. Expression of selected adipose genes in preparations of bone marrow cells, adipocytes isolated from bone marrow and adipocytes isolated from epididymal adipose tissue. A set of original RNA from the same animal was re-amplified to aRNA, then converted to cDNA. Relative fold change was normalized to endogenous 18S and bone marrow stromal cells. Data are presented as Log2 of fold change.