The knowledge reliability was measured employing the scatter plot technique. The correlation coefficients (Rs) of the normalized samples ranged from .ninety five to .ninety nine, which signifies that a systematic KIN1408 chemical information variation of non-organic origin was taken off. Comparative evaluation amongst the motor vehicle-handled rats and AEtLP-treated rats samples was executed using a t-examination (|fold|> or <2) and the adjusted Benjamini-Hochberg FDR (false discovery rate) (P <0.05). Also, if the value of fold change are between 0.5< and <1.5 after AEtLP treatment, these gene decided as recovered genes and now functioning normally. Furthermore, volcano plots and hierarchical cluster analysis were conducted using a complete linkage and Euclidean distance as a measure of similarity. Time-dependent profiling was achieved by k-means cluster analysis (k 9, Euclidean distance, complete linkage). All data analysis and visualization of the differentially expressed genes was conducted using ArrayAssist (Stratagene, La Jolla, USA). The biological pathway and ontology-based analysis were performed using the PANTHER (Protein ANalysis THrough Evolutionary Relationships) database (http://www.pantherdb.org). All microarray data was submitted to the GEO database (http://www.ncbi.nih.gov/geo) under accession number GSE62041.The complementary DNA was synthesized using mixture containing the total RNA (5 g), oligo-dT primer (500 ng, Invitrogen, CA, USA), dNTP and reverse transcriptase (200 units, Invitrogen) according to the method described elsewhere [76]. Furthermore, the PCR product of each gene was amplified using the specific primers, as described in previous studies [77].Firstly, in order to separately collect muscle and mucosa layer, the distal colon of SD rats were cut along a colon tube using scissors and washed with 1x PBS. Next, the mucosa layer were scraped out from the muscle layer using surgical knife and collected in homogenized tube. Whole distal colon, muscle layer and mucosa layer collected from a subset of the groups were homogenized using a PRO-PREP Solution Kit (iNtRON Biotechnology, Sungnam, Korea) supplemented with half of a protein inhibitor cocktail tablet (Roche, Penzberg, Germany), then centrifuged at 10,000 for 10 min. The prepared proteins were subsequently subjected to 10% SDS-PAGE, after which they were transferred to a nitrocellulose membrane (Amersham Biosciences, Corston, UK) for 2 h at 45 V in transfer buffer (25 mM Trizma-base, 192 mM glycine, and 20% methanol). The efficiency of the transfer and equal protein loading were evaluated by staining the membrane with Amido Black Staining Solution (Sigma-Aldrich Co.) and the gel with Coomassie Blue. Appropriate dilutions of primary antibodies, anti-FGF antibody (Santa Cruz, CA, USA), anti-ACE antibody (R&D Systems, Minneapolis, MN, USA) and anti-actin (Sigma-Aldrich Co.) were added to the membranes and allowed to hybridize overnight at 4. After the antibodies were removed, the membrane was washed three times in a solution composed of 10 mM Trizma-base (pH 7.6), 150 mM NaCl, and 0.05% Tween-20 for 10 min. The membrane was then incubated with horseradish peroxidase-conjugated anti-secondary antibody for 1 h at room temperature, after which it was washed again as described above and developed using an enhanced chemiluminescence detection system (Amersham Biosciences). Finally, the results were quantified using the Image Analyzer System (Eastman Kodak 2000MM, Eastman Kodak, Rochester, NY, USA) and expressed as1654254 the fold-increase over control values. These results were confirmed by two independent researchers conducting the experiments at 
least twice.
