Another recognized limitation of our study is the small sample size in the peptide group. To determine the extent to which this limitation impacts the ability to draw conclusions from the data power analysis and sample size calculations were performed. The sample size calculation was based on the observed standard deviation of 5.54% for the percent of total arch that contains lesion and an observed difference in the means of 2%. Considering these statistics and a desired power. 80%, a sample size of greater than 100 per group would be needed to identity significant differences. Based on these sample size calculations it is not expected that doubling or tripling the sample size of the current study would result in significant differences. Possible contributors to reduced sensitivity to peptide in this model may be not only the absence of the LDLR but also the interaction with the highly obesogenic diet. With respect to adiposity and glucose tolerance, one study used chow-fed ob/ob mice injected with L4F at the dose of 2 mg/kg/ day and demonstrated attenuated obesity and adipose inflammation. In that study, L4F treatment also improved both glucose and Niraparib insulin tolerance tests. While the ob/ob model consistently develops obesity related insulin resistance, the hyperlipidemia is characterized by elevations in HDL and this does not associate with atherosclerosis development. We are the first to have reported effects on weight gain, inflammation, insulin resistance, and atherosclerosis in a model of DIO with lipid profiles similar to humans. Our ability to compare the effects of L4F treatment directly to that of apoA-I overexpression is another strength of this research. We have shown using the HFHSC diet in C57BL/6 mice that were either wild type or transgenic with respect to apoA-I expression, apoA-I overexpression reduced adipose tissue inflammation. Similar reductions of adipose tissue inflammation by apoA-I overexpression were also seen in the Ldlr background, with no change in total plasma cholesterol or triglyceride levels. It is generally thought that adipose tissue inflammation is related to several systemic perturbations, such as glucose intolerance. In the present study, despite the reduction of adipose tissue inflammation, apoA-I overexpression did not influence weight gain or glucose sensitivity in the DIO Ldlr male model. This suggests that the gain of adipose tissue mass is not strictly correlated with the degree of inflammation in this tissue. These data argue that the relationship between adiposity, weight gain, adipose tissue inflammation, and glucose intolerance is more complex than has been thought and are not directly correlated. Peptide treatment of Ldlr mice on the HFHSC diet also did not influence glucose tolerance at a moderate dose of 100 mg/day/mouse. Peptide treatment could have two major actions: influence on cholesterol homeostasis and reduced oxidized lipid concentrations. In our model, there is no evidence of these actions of peptide being operative in vivo. Perhaps the drive of our dietary model toward atherogenesis and glucose intolerance is so strong as to be beyond the reach of the peptide at the level we employed, which was dosed based on studies in apoE and ob/ob mice. It is not clear whether obesity in this model would indeed be responsive to higher doses of peptide. While a majority of the studies have been conducted in apoE mice, this model is driven by hyperlipidemia and does not lend itself to investigation of several other aspects of the metabolic syndrome, namely DIO and insulin resistance.