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Bacterial Colony-forming units (CFU) assay

YF Yesheng Fu
LL Lei Li
XZ Xin Zhang
ZD Zhikang Deng
YW Ying Wu
WC Wenzhe Chen
YL Yuchen Liu
SH Shan He
JW Jian Wang
YX Yuping Xie
ZT Zhiwei Tu
YL Yadi Lyu
YW Yange Wei
SW Shujie Wang
CC Chun-Ping Cui
CL Cui Hua Liu
LZ Lingqiang Zhang
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Cells were seeded in antibiotic-free complete medium. The following day, cells were infected with S. flexneri (MOI ~ 100), the bacteria were washed off with PBS 2 h post infection and placed in fresh DMEM containing 50 μg/ml gentamycin (to kill extracellular bacteria). After another 2 h of incubation, infected cells were washed three times with PBS, and placed in medium containing 10% FBS. After 12 h of infection, the attached and detached cells were collected and disrupted by the addition of sterile water with 1% Triton X-100, diluted, and plated onto agar plates and the number of internalized bacteria was determined by counting the colony-forming units after incubation at 37 °C.

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