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Measurement of adrenal stress response In a separate study, WT, hl?/? hHL and hl?/? ciHL mice on HFD were subjected to a 16?h fast and blood withdrawn with minimal handling ( was used on pooled data from male and female mice to compare energy expenditure between genotypes after adjusting for differences in body mass VE-821 ic50 and composition. Statistical analyses of energy expenditure were performed by the NORC Biostatistics Subcore using SPSS (version 19, IBM Corp., Somers, NY). No statistical analysis was performed for the historical data on WT mice. Results Expression of hHL and ciHL Expression of active and inactive human HL in vivo was confirmed by analysis of postheparin plasma and demonstrated physiologic HL activity (in the range of human HL activity) in hl?/? hHL mice but not in either hl?/? mice that lack HL or in hl?/? ciHL mice rescued with the catalytically inactive transgene (Table?(Table2).2). As expected, HL activity was present in preheparin plasma from WT learn more mice, reflecting the reduced heparin affinity of murine HL that allows a large portion of HL to circulate (Table?(Table2).2). Background activity from incompletely suppressed lipoprotein lipase was detected in pre-heparin plasma from hl?/?, hl?/? ciHL, and hl?/? hHL mice as well as in postheparin plasma from hl?/? and hl?/? ciHL mice (Table?(Table2).2). Western blot analysis confirmed the presence of immunoreactive human HL in both hl?/? hHL and hl?/? ciHL mice and its absence in hl?/? and WT mice (Fig. 1). Figure 1 Western blot analysis for human HL immunoreactivity. Expression of human HL in postheparin plasma from hl?/? ciHL (Left Azastene panel) and hl?/? hHL mice (Right panel) were assayed by Western blot using a monospecific antihuman ... Table 2 Hepatic lipase activities in pre and postheparin plasma1. Genotype differences in body weight and body fat mass Body weight at baseline was similar among the three genotypes, HL-deficient mice alone or expressing either intact hHL or catalytically inactive ciHL (Table?(Table3).3). However, following exposure to HFD for three months, male hl?/? mice rescued with transgenic expression of intact hHL (possessing both the catalytic and bridging functions) had significantly higher body weight (?15%) than either male mice rescued with inactive ciHL (which contains the bridging function, but is catalytically inactive) or hl?/? male mice alone (P?