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. 2017 Oct 24;38(6):3347–3354. doi: 10.3892/or.2017.6056

Figure 2.

Figure 2.

Knockdown of lncRNA-XIST inhibits basal autophagy and autophagic flux in NSCLC cells. (A) Efficiency of shRNA sequences against lncRNA-XIST was examined by quantitative real-time PCR (qPCR) both in A549 and H1299 cells. Data are shown as the mean ± SD of three independent experiments. (B) A549 and H1299 cells were transfected with shRNA-lncRNA-XIST (Sh-X) or the control plasmid (Ctrl). After 72 h, the expression of SQSTM1, LC3BI, LC3BII and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were detected by western blotting in the cells. (C and D) A549 and H1299 cells were co-transfected with shRNA-lncRNA-XIST (Sh-X) and GFP-LC3B plasmid or the control plasmid (Ctrl) for 72 h, and the number of GFP-LC3B dots were examined by (C) confocal microscopy (D) and quantified for each cell line. Data are showed as mean ± SD of three independent experiments. (E) A549 and H1299 cells were transfected with Sh-X or Ctrl for 72 h and treated with or without 25 µM CQ for 4 h. The expression of LC3B-II and SQSTM1 was examined by western blotting. (F) The autophagic flux was shown by the quantitation of the rate of LC3BII/GAPDH in cells cultured with 25 µM CQ. Data are shown as mean ± SD of three independent experiments. *P<0.05; **P<0.01; ***P<0.001.