Fig. 4

FOXA2 identified by PCR array was differentially expressed between spheres and parental cells and regulated by autophagy. (a) GO term enrichment analysis indicated that the 26 genes differentially expressed (more than 3-fold) could be categorized into cancer stem cell markers, loss of stemness, therapeutic targets, signal pathways, pluripotency, asymmetric division and migration. (b)Expression of 10 selected genes was validated between 3AO, SKOV3 parental and sphere cells by qRT-PCR. (c)Alteration of the 10 selected genes was determined using qRT-PCR after autophagy was inhibited using BafA1 in 3AO and SKOV3 sphere cells. (d) Protein levels of FOXA2, ALDH1A1, and IL8 were determined by Western Blotting in 3AO and SKOV3 parental and sphere cells treated with BafA1(10 nM, 48 h) or not. GAPDH was analyzed as the loading control. (e)The mRNA and protein levels of FOXA2 were measured in 3AO and SKOV3 spheres after autophagy was inhibited by ATG5 knockdown or CQ(10 μM, 48 h).Three independent experiments were performed and the results were expressed as the means ± SD, and analyzed using Student’s t-test (**P < 0.01)