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Fig. 2 | Molecular Cancer

Fig. 2

From: Blockade of autophagy reduces pancreatic cancer stem cell activity and potentiates the tumoricidal effect of gemcitabine

Fig. 2

Blockade of autophagy by genetic knockdown reduces pancreatic CSC activity. a The expression of autophagy-related proteins and CSC markers in the PANC-1-derived cells permanently expressing shRNAs against ATG5 (shATG5), ATG7 (shATG7), and BECN1 (shBECN1) was detected by Western blotting. The levels of LC3-II, ALDH1, CD133, and CD44, were normalized against β-Actin and presented as means ± SE of three separate experiments. NS, not significant; ***, P < 0.001, vs. control cells. b The expression of LC3, ALDH1, CD44, and CD133 in the control, shATG5, shATG7, and shBECN1 cells was analyzed by flow cytometry. The bar graphs indicate the percentage ± SE of LC3+/ALDH1+ cells (left) and CD44+/CD133+ cells (right). c The enzymatic activity of ALDH1 in the control, shATG5, shATG7, and shBECN1 cells was examined by ALDEFLUOR assay. The bar graph indicates the percentage ± SE of ALDEFLUOR+ cells in each cell line. d The bright field micrographs showed sphere formation of the control, shATG5, shATG7, shBECN1 cells cultured in ultra-low attachment plates for 14 days. The number of spheres was calculated and presented as means ± SE. NS, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001 compared to controls

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