Quantity one image analysis software

For quantification, developed films were scanned at 2400 × 2400 dpi (i800 MICROTEK high-quality film scanner), and the densitometric analysis was performed using Quantity One Image software analysis (Biorad, Windows) and the Gel-Pro analyzer (Media Cybernetics, Windows). Statistical analysis of the obtained data was performed using Bonferroni post hoc test (Multiple comparison test) using GraphPad Prism 5.0c (Mac OsX, Grahpad). Data are presented as mean ± standard error of the mean (S.E.M.). Differences between groups were considered statistically significant at *P < 0.05.

The meta-genomic DNA was extracted from the frozen faecal content of rats by the QIAamp DNA stool mini kit (Qiagen, Hilden, Germany). PCR was conducted using universal primers F338+GC clamp and R518 targeting the hyper-variable V3 region of 16S rRNA gene. The resulting 16S rDNA amplicons were analysed using the DCode system (Bio-Rad, Hercules, CA, USA) according to descriptions of Joossens et al. (2011) . The digitised denaturing gradient gel electrophoresis (DGGE) images were analysed with Quantity One image analysis software (version 4.6.1; Bio-Rad). Similarities were displayed graphically as a dendrogram.

The level of focal adhesion development was semi-quantitatively examined in total protein extracts and separated protein fractions (S and C) by Western blotting for b1-integrin and vinculin. Equivalents of 5 mg total protein or 12.5 mL of the separated protein fractions were size fractioned on a 10% polyacrylamide gel and blotted onto a Hybond PVDF membrane (Immobilon-P, Millipore). Non-specific binding sites were blocked with 5% milk powder (ELK) in 0.1% PBS/Tween20. Membranes were probed with primary antibodies at 4 1C overnight, washed and subsequently incubated at room temperature with horse-radish peroxidase (HRP) labeled secondary antibodies for 1 h (for used antibodies and their dilutions see Table 1). For total protein extracts, GAPDH was used as loading control. For visualization, membranes were incubated with enhanced chemiluminescence substrate (Thermo Scientific Pierce), and bands were detected using the Proxima C16 Phi+ (Isogen) imager. Bands were quantified by determining the intensity of the bands (INT*mm 2 with global background correction) using Quantity One image analysis software (Biorad, version 4.6.6).

An aliquot of 0.1 mg of hatchlings TL was taken for determine the esterification pattern of [1-14 C]ARA into the different LC. Lipid classes were separated by one-dimensional double-development HPTLC as previously described 2.4. Developed HPTLC plates were placed for 2 weeks in closed exposure cassettes (Exposure Cassete-K, BioRad, Madrid, Spain) in contact with a radioactive-sensitive phosphorus screen (Imagen Screen-K, Biorad, Madrid, Spain). The screens were then scanned with Molecular Imager FX image acquisition system (BioRad, Madrid, Spain), and the bands were quantified using the Quantity One image analysis software (BioRad, Madrid, Spain). Identification of labelled bands was confirmed by radiolabelled standards simultaneously run on the same plate (Rodríguez et al., 2002) . andL-∝-1-palmitoyl-2-[1-14 C