Figure 5. mTOR signaling pathway and autophagy related proteins detection. (A) Bioinformatics prediction (http://www.string-db.org):mTOR and UPF1 maybe co-expression and interaction. However, mTOR and ULK1 show that both of its relationship have verified. (B, C) Transfected with pcDNA3.1-UPF1+ and pcDNA3.1 or siUPF1 and NC in Ishikawa showed that UPF1 could affect mTOR and downstream’s proteins. (D, E) Overexpressed UPF1 or downregulated UPF1 could inhibit or promote autophagy. LC3I/II and p62 showed autophagy weakened or increased. (F–I) The immunofluorescence assays were performed in EEC cells that were transfected with mRFP-GFP-LC3 lentiviral in two different groups. The numbers of GFP and mRFP dots were determined by fluorescent puncta in three high-power fields. The statistical significance between different groups. Data was represented as the mean +/- SEM.*P<0.05, **P<0.01, ***P<0.001.