Figure 7. Quantitative analysis of shows that bezafibrate significantly increases nuclear distribution of L-FABP in cultured mouse hepatocytes.

Panels A and B: WT mouse hepatocytes were cultured overnight, washed, and further incubated for 0.5–24 h with serum-free, glucose-free Williams’ medium E plus 6 mM (A) or 20 mM (B) glucose, insulin, dexamethasone, and LCFA-free BSA (Alb) or BSA/bezafibrate (200 μM) as described in Methods. Cells were fixed, labeled with FITC-anti L-FABP and TO-PRO DNA dye, and multiple cells analyzed by confocal microscopy to determine nuclear and cytoplasmic L-FABP labeling intensity in order to calculate the nuclear/cytoplasm L-FABP ratio (Nucl/Cyto FI L-FABP) described in Methods [29,30,117]. Means ± SEM, n = 40. *p < 0.05 vs albumin; # p < 0.05 as compared at 0.5 h within each group. C: WT hepatocytes were cultured as above and incubated with 6mM glucose and either LCFA-free BSA or BSA/bezafibrate (200 μM) for 24 hrs, fixed, anti-L-FABP immunogold labeled, and antibody-L-FABP labeling particle density in the whole cell, nucleoplasm and cytoplasm determined as we described [29]. Statistics was performed using a one-way ANOVA and a Newman-Keuls post-test. Mean ± SEM, n = 20 @ p<0.05 vs Alb whole cell; # p<0.05 vs Alb nucleus; $ p<0.05 vs Alb cyto; ^ p<0.05 vs BZ whole cell; & p<0.05 vs BZ nucleus.