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Cell staining and fixation

GX Gongyu Xu
HS Hao Shen
EN Emile Nibona
KW Kongyue Wu
XK Xiaomei Ke
MH Md. Abdullah Al Hafiz
XL Xiaoting Liang
XZ Xueping Zhong
QZ Qingchun Zhou
CQ Chao Qi
HZ Haobin Zhao
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In order to label mitochondria, GCO and 239 T cells were incubated with pre-warmed (37 °C) MitoTracker Green FM probes (Thermo Fisher Scientific; M7514) at a concentration of 200 nmol/L for 40 min under growth conditions in the dark. Nuclei were stained with Hoechst 33258 (Thermo Fisher Scientific; H3569). After staining, the solution was replaced with a fresh pre-warmed culture medium. Images were taken using an EVOS FL auto fluorescence microscope (Thermo Fisher Scientific).

In order to label lysosomes, CellLight Lysosomes-GFP reagent (Thermo Fisher Scientific; C10596) was added to cells in a complete culture medium, following the manufacturer’s instructions, and was gently mixed. Cells were incubated with the reagent overnight (≥16 h) in the dark. Images were taken with a confocal microscope (Leica, Germany; SP5) following cell fixation.

Copyright and License information: The Author(s) ©2019
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