Figure 2.

CU27 diminishes liver CSC‐associated phenotypes and functions in vitro. A,B) Representative immunofluorescence images and flow cytometry analysis of expression levels of CK18, CK19, and AFP besides CD13 in CU27‐treated HCC cells (10 µm, 48 h, n = 3). Bars, 100 µm. C) Representative images of untreated HCC cells‐derived tumorspheres simultaneously treated with CU27 (7 days). Bars, 1000 µm. D) Representative images and quantitative analysis of tumorspheres from sorted CD13⁺/CD133⁺/EpCAM⁺ HCC cells after a pre‐treatment with CU27 (48 h) in CDM culture before proceeding to tumorsphere formation. Bars, 1000 µm. Mean ± SD, ^** p < 0.01, data analyzed by Student's t‐test. E,F) Chemoresistance assay of doxorubicin (Dox), sorafenib, and 5‐Fluorouracil (5‐FU) in sorted CD13⁺/CD133⁺/EpCAM⁺ HCC cells treated with CU27 (0.5 µm) in CDM culture (IC₂₀ of CU27, 48 h, n = 3). IC₂₀, 20% growth inhibition concentration of CU27; Means ± SD; ^* p < 0.05, ^** p < 0.01; two‐way ANOVA.