Figure 5

Exogenous addition of LPL or palmitate reverses the effect of LPL-siRNA treatment on lipid accumulation by adipocytes. 3T3-L1 cells were transfected with or without LPL-specific siRNA (100 nM), followed by incubation with adipocyte induction reagents. In parallel, some cells were also incubated with palmitate-albumin (2 mg/ml protein) beginning with the second addition of insulin during the differentiation protocol. Alternatively, LPL (6.67 Units/ml) was added with the second addition of insulin and each successive day until measuring intracellular lipid levels. After a total of five days, intracellular lipid accumulation was quantified by measuring cell-associated Nile Red using fluorometry.