MKN45 cells or MKN45 cells pretreated with H. pylori for 12 h were plated in 24-well culture plates that contained two layers of soft agar (Corning, USA) at a density of 500 cells/well and allowed to grow undisturbed for 14 days, at 37 °C, high humidity, and 5% CO2. The top and bottom layers were 0.33% and 0.8% low melt agarose (Bio-Rad, Hercules, CA, USA) in 5% RPMI 1640 medium, respectively. Colonies were counted by two investigators (QJ and XL).
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