Although human liver fatty acid binding protein (FABP1) T94A variant has been associated with non-alcoholic fatty liver disease (NAFLD) and reduced ability of fenofibrate to lower serum triglycerides (TG) to target levels, molecular events leading to this phenotype are poorly understood. Cultured primary hepatocytes from female human subjects expressing the FABP1 T94A variant exhibited increased neutral lipid (TG, cholesteryl ester) accumulation associated with: 1) upregulation of total FABP1, a key protein stimulating GPAM the rate limiting enzyme in lipogenesis; 2) increased mRNA expression of key enzymes in lipogenesis (GPAM, LPIN2) in heterozygotes; 3) decreased mRNA expression of microsomal triglyceride transfer protein (MTTP); 4) increased secretion of ApoB100 but not TG; 5) decreased LCFA β-oxidation. TG accumulation was not due to any increase in long chain fatty acid (LCFA) uptake, de novo lipogenesis, or the alternate MOGAT pathway in lipogenesis. Despite increased expression of total FABP1 mRNA and protein, fenofibrate mediated FABP1 redistribution to nuclei and ligand-induced PPARα transcription of LCFA β-oxidative enzymes (CPT1A, CPT2, and ACOX1) were attenuated in FABP1 T94A hepatocytes. While the phenotype of FABP1 T94A variant human hepatocytes exhibits some similarities to that of FABP1 null or PPARα null hepatocytes and mice, expression of FABP1 T94A variant did not abolish or reduce ligand binding. Thus, the FABP1 T94A variant represents an altered/reduced function mutation resulting in TG accumulation.