G box xr5 gel imaging system

Different amount of OVA and BSA were first separated in a similar gel by SDS PAGE. One gel was stained with Coomassie brilliant blue R250 solution in (50% ethanol and 10% acetic acid in water), distained with distaining buffer (50% ethanol and 10% acetic acid in water) and photographed in G:BOX XR5 Gel imaging system (Syngene, Cambridge, UK). Pro-Q^® Diamond gel staining was done according to the user manual. In brief, the gel was fixed in 50% methanol plus 10% acetic acid for 30 min twice, washed 3 times with ultrapure water each for 10 min, stained with 60 ml Pro-Q^® Diamond stain for 90 min, distained with 20% acetonitrile in 50 mM sodium acetate pH4.0, wash twice with ultrapure water and imaged with Typhoon FLA 9000 (GE Healthcare Waukesha, WI, US).

Transfected HCT116 and RKO cells were plated in 6-well plates at a density of 800 cells in triplicate and incubated at 37 °C with 5% CO₂ for 7–10 days until they reached 80% confluence. Cells were then fixed with 4% paraformaldehyde and stained with 0.5% crystal violet (Sigma Aldrich; Merck KGaA) for 30 min at room temperature. Images were captured on a G: BOX XR5 Gel Imaging System (Syngene, UK). After dissolving in DMSO, colonies were quantified and the absorbance was measured on a microplate reader (MD, USA) at 570 nm.