Fig. 1

Identifying CSC like properties in Hepa1–6 spheroids: a Hepa1–6 adherent cells (left, control) and 7 days old spheroids (right, serum-free); b Flow cytometry data showing significant higher EpCAM expression in spheroids compared with control (Day 7, **p < 0.005, n = 4); c ICC staining for EpCAM (FITC - green) confirming flow cytometric findings that EpCAM upregulated in spheroids. Nuclear stained with DAPI (blue); d CSCs spheroids also showed significant increases in CD44 and CD90 compared with control (p < 0.05, n = 3). e Hoechst efflux assay: Hepa1–6 spheroids showed significantly higher ABCG2 expression by western blot (upper panel) and significantly increased Hoechst-33342 dye efflux activity (lower panel). SF = Hepa1–6 spheroids (n = 3 independent experiments)