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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: Autophagy maintains the stemness of ovarian cancer stem cells by FOXA2.

Fig. 2

.The autophagic flux was enhanced in OCSCs. (a) Protein levels of LC3B, SQSTM1 and ATG5 were detected by Western Blotting in 3AO/SKOV3 parental and sphere cells. (b) Protein levels of SQSTM1 and LC3B were detected by Western Blotting in 3AO and SKOV3 parental and sphere cells. Cells were cultivated in complete medium (Ctrl) or starvation conditions (induced with EBSS for 4 h) in the presence or absence of BafA1 (final concentration 50 nM) for 4 h. GAPDH was analyzed as the loading control. The LC3B-II/GAPDH ratio was determined using the Quantity One software. The autophagic flux was determined as the ratio between the LC3B-II levels with and without BafA1 in histograms (right). (c)3AO-GFP-LC3B and SKOV3-GFP-LC3B parentaland sphere cells were cultivated as above, then visualized by confocal fluorescence microscopy. The number of GFP-LC3B-II positive puncta in 50 GFP-positive cells for each group was counted using the Metamorph offline 7.7.8.0 software package and the ratio between the GFP-LC3B-II-positive puncta per cell with BafA1 and without BafA1 was calculated (right). Three independent experiments were performed and the results were expressed as the means ± SD, and analyzed using Student’s t-test (* P < 0.05, **P < 0.01)

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