A549 and H1975 cells (1000 cells per plate) were suspended in 1 mL of DMEM or RPMI 1640 containing 0.3% low-melting-point agarose (Amresco, USA), 10% FBS and indicated concentration of HHT, andplated on a bottom layer containing 0.6% agarose and 10% FBS in 6-well plate intriplicate. After 2 weeks, plates were stained with 0.5 mL of 0.005% crystal violet for more than 1 h and the colonies were counted under light microscope60 (link).
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