Chronic glucolipotoxic conditions in vitro impair fatty acid uptake/ metabolism. Rat islets and NIT1 cells were cultured under normal conditions (control) or with 16.7 mM glucose and 500 μM palmitate for 72 h (GL). Post 72 h, RNA was isolated from islets and the mRNA levels of cd36 (A) and PPARα (E) were measured as described in the materials and methods. Protein levels of CD36 in both islets and NIT-1 cells (B) were measured by western blotting using anti-CD36 antibodies. Fatty acid uptake (C) was measured in NIT-1 cells using green-fluorescent BODIPY dyes, a non-metabolized fluorescent labelled fatty acid analog as described in the materials and methods. For triglyceride estimation, post incubation cells were lysed and cellular triglyceride levels were estimated and normalized to total protein content (D). Data are expressed as mean±SEM and statistical analysis was performed using the unpaired Student’s t-test. (*P<0.05, **P<0.01 and ***P<0.001, n=4).
Somesh et al. BMC Cell Biology 2013 14:31 doi:10.1186/1471-2121-14-31