9- and 4.0-fold, respectively) but unaffected by TFA alone (Table 3; P < 0.05). Ketogenic Fibroblast Growth Factor 21, a key mediator of hepatic lipid mobilization, was induced 2.4-fold. Additionally, expression of genes associated with hepatic fibrosis, including ��-laminin (Lamb3b) and ��-actin (Actg1), selleck chem Sorafenib were increased in the TFA+MSG livers by 1.9- and 1.5�Cfold, respectively (Table 3; P �� 0.05). The MSG diet induced an increase in the expression of a number of key transcriptional regulatory proteins, including Growth Arrest and DNA damage-inducible 45B (GADD45b; 2.7-fold) and NIPA-like domain containing 1 (2.3-fold); in each case, expression returned to control levels in animals treated with the TFA+MSG diet. The TFA+MSG diet also induced the expression of genes involved in the inflammatory pathway, including a 1.

4-fold increase in Interferon-�� 14, a 7.5-fold increase in adipsin (complement factor D), and a 2-fold increase in defensin ��1. Hepatic expression of apoptotic regulator p21 was increased by 2.4-fold in the TFA+MSG diet, further suggestive of a link between oxidative stress, inflammation, and cell cycle impairment in this model of diet-induced NAFLD. Confirmation of Affymetrix microarray by real-time qRT-PCR A set of five genes was randomly selected for validation via real-time qRT-PCR using mRNA derived from mice other than those used for the microarray study (n = 4 per diet group).

Figure 3A shows qRT-PCR analysis of hepatic expression of CIDEC (“type”:”entrez-nucleotide”,”attrs”:”text”:”NM_178373″,”term_id”:”141802598″,”term_text”:”NM_178373″NM_178373), cyp7a1 (NM_ 007824), GADD45b (“type”:”entrez-nucleotide”,”attrs”:”text”:”NM_008655″,”term_id”:”6678979″,”term_text”:”NM_008655″NM_008655), MTTP (“type”:”entrez-nucleotide”,”attrs”:”text”:”NM_008642″,”term_id”:”254540222″,”term_text”:”NM_008642″NM_008642), AV-951 and SREBP1c (“type”:”entrez-nucleotide”,”attrs”:”text”:”NM_011480″,”term_id”:”146134488″,”term_text”:”NM_011480″NM_011480) levels relative to housekeeping ��-actin levels in mice from the four different diet groups (n = 4, P < 0.05). Figure 3B shows representative ethidium bromide-stained RT-PCR products electrophoresed on 2% agarose gels. Figure 3C shows concordance of qRT-PCR results versus microarray results expressed as signal intensity (percentage control diet �� SD). Pearson correlation coefficients (r) are indicated on each graph. Values of r were >0.9 in all cases, suggesting a high level of agreement between the microarray and the qRT-PCR data. Individual values were as follows: CIDEC, 0.99; Cyp7a1, 0.99; GADD45b, 0.99; MTTP, 0.99; and SREBP1c, 0.96 (P < 0.05). Fig. 3. Validation of Affymetrix microarray analysis of gene expression using qRT-PCR.