Figure 2. Screening of natural compounds that selectively impede the stemness of iCSCL-10A cells.

(A) A list of natural compounds assayed in the study. (B, C) iCSCL-10A cells (5×10³ cells/well in 100 μl) were plated in 96-well plates. After 24 hrs, 1 μM of compound was added to the appropriate wells. Cell viability was measured after 48 hrs using WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium) activity assays (B). Alkaline Phosphatase (ALP) activity was measured after 48 hrs using the TRACP & ALP Assay Kit and a microplate reader. Values represent the mean ± SEM (n = 3) (C). (D) Quantification of tumor sphere formation. iCSCL-10A cells were plated on ultra-low attachment surface 96-well plates in the presence of 1μM of the compounds and tumor sphere formation was assessed at 48 hrs.