Soft agar colony formation assay

XL Xuan Liu
QJ Qing Ji
CZ Chengcheng Zhang
XL Xiaowei Liu
YL Yanna Liu
NL Ningning Liu
HS Hua Sui
LZ Lihong Zhou
SW Songpo Wang
QL Qi Li
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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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