Figure 3: Fat tissue filtrate from mice fed a high fat diet stimulates the proliferation of JB6 cells growing in liquid medium. A. JB6 cells were treated with 1% FBS, 5% FBS or fat tissue filtrate (50 μg protein/mL) from mice fed a HFD for 4 months and were counted at 1, 3, 5, and 7 days. The fat tissue filtrate from mice fed a HFD stimulated JB6 cell proliferation to about the same extent as 5% FBS. One well = 9.6 cm2. B. JB6 cells were seeded at 100 cells per well in a 6-well plate. Cells were treated with 5% FBS or fat tissue filtrate from mice fed a HFD for 7 days and stained with Giemsa. Cloning efficiency, as measured by the number of Giemsa-stained clones, was significantly increased to a greater extent in JB6 cells cultured with fat tissue filtrate from mice fed a HFD for 4 months compared with cells cultured with fat tissue filtrate from mice fed a LFD for 4 months (*P<0.05). Arrows indicate % increase in cloning efficiency at the different doses. One well = 9.6 cm2. Data are mean ±SEM, Student t test (unpaired, two-tailed).