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. 2017 Sep 18;7:11776. doi: 10.1038/s41598-017-10407-w

Figure 6.

Figure 6

miR-486 directly regulate post-transcriptional expression of BCAP. (A,B) A549 and LLC cells were treated as described in Fig. 2C. Expression levels of BCAP were detected by qRT-PCR. (C) Luciferase reporters containing wild-type (WT) or mutant (MUT) miR-486 binding sites in the BCAP 3′-UTR were co-transfected into 293 T cells along with control luciferase reporter plasmid, miR-NC, miR-486 mimic for 48 hours. (D–E) qRT-PCR analysis of BCAP mRNA levels and Western blot analysis of BCAP protein levels in A549 cell treated with miR-486 mimic. (F) A549 and LLC cells were co-transfected with GFP-LC3 plasmid, siNC or siBCAP for 48 h. Number of autophagosomes was quantified. (G) A549 and LLC cells were transfected with GFP-LC3 plasmid, miR-NC, miR-486 mimic or co-transfected with miR-486 mimic and BCAP plasmid. Number of autophagosomes was quantified. (H) A549 and LLC cells transfected with siNC or siBCAP for 48 h. Western blot analysis of phospho-AKT, AKT, phospho-mTOR, mTOR and GAPDH expression. (I) A549 or LLC cells transfected with miR-NC, miR-486 mimic or co-transfected with miR-486 and BCAP plasmid. Western blot analysis of phospho-AKT, AKT, phospho-mTOR, mTOR and GAPDH expression. All error bars indicate mean ± SEM. Experiments were repeated three times independently. **P < 0.01.