Powershot a640

Human monocyte leukemic cell line (THP-1) was purchased from (ATCC, UK). Nonadherent cells were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS (Biowest, France), 2 mM l-glutamine and 100 U/mL streptomycin-penicillin (Biowest, France) at 37 °C with 5% CO₂. Foam cell formation assay was conducted as described by Khan et al. (2014) [41 (link)]. Briefly, THP-1 cells plated at 3 × 10⁵ cells/well on glass coverslips (placed in 12 well plate) were differentiated into macrophages with 50 nM phorbol-12- myristate-13-acetate (PMA; Sigma-Aldrich) for 72 h at 37 °C with 5% CO₂. After 72 h, the adherent macrophages were exposed to H. pylori GE (10 ug/mL, 1 ug/mL, 0.1 ug/mL), H. pylori LPS (25 ng/mL and 5 ng/mL), LPS E. coli (25 ng/mL) and 7-KCh (conc. 40 ng/mL) for 24 h at 37 °C with 5% CO₂. After 24 h, the cells were fixed with 4% paraformaldehyde in PBS for 15 min, and covered with Oil Red O (Sigma) at a working solution (0.6% (m/v) in a 60% (v/v) isopropanol solution for 30 min. Next, the cells were counterstained with a hematoxylin solution (Elektro Med, Niepolomice, Poland) for 3 min. Stained cells were visualized with light microscopy (Eclipse 50i, Nikon; 1000× magnification) with a photo camera, a Power Shot A 640 (Canon), integrated. Lipid droplets appeared red and nuclei appeared blue.

The aforementioned constructs harboring 35S::FaTHSFA2a, 35S::FaTHSFA2aΔAD, 35S::FaTHSFB1a, or 35S::FaTHSFB1aΔRD were transformed into wild-type Arabidopsis plants (ecotype: Col-0) by using the floral-dipping method [69 (link)]. The seeds were harvested and surface-sterilized with 25% sodium hypochlorite solution. These transgenic plants were selected on solid half MS medium containing 30 μg/mL hygromycin. The plates were sealed with 3 M micropore tape and placed vertically in a growth chamber (CH-202, CHIN-HSIN, Taipei, Taiwan), with a long-day condition (16-h light/8-h dark cycle), and 120 Lmol·m⁻²·s⁻¹ photo flux density at 22 ± 2 °C for 12 days before transplanting to soil. The ectopic gene expression of FaTHSFA2a, FaTHSFA2aΔAD, FaTHSFB1a, and FaTHSFB1aΔRD was confirmed through RT-PCR by using gene-specific primer sets (Table S4). Each transgenic whole plant was photographed using a digital camera (Canon, PowerShot A640, Tokyo, Japan). Young seedlings of wild-type and all transgenic plants were viewed under a dissecting microscope (Leica, MS5, Heerbrugg, Switzerland), and all image data were collected using a digital camera (Canon, PowerShot S80, Tokyo, Japan) under a light-field.

The collagen proportionate area (CPA) was measured as described previously [18 (link)]. Briefly, image capture was accomplished with a Canon Powershot A640 digital camera and digital image analysis (DIA) used a visual basic script for Zeiss Axiovision (version 4.8.2., Carl Zeiss Ltd., Cambridge, UK). The algorithm uses a binary segmentation of RGB color channels to distinguish liver tissue from collagen and an editing step was included. This allows manual editing of confounding artefacts such as major blood vessels and liver capsule. The collagen proportionate area was calculated as the area occupied by the collagen as a proportion of the area of the whole parenchyma and expressed as a percentage [18 (link)].