Fig 2: Knockdown and overexpression of miR-199a-5p affects cellular autophagy levels. (A) Western blot analysis of LC3I and LC3II protein expression levels in various cell lines and the LC3II/LC3I ratio with or without CQ treatment. β-actin was used as the loading control. (B) Neutral red staining for lysosome analysis. Scale bar, 10 µm. (C) Western blot analysis of the protein levels of PI3K/Akt/mTOR pathway, eEF2K, p62 and ATG5 proteins and the quantitative analysis results. The experiments were repeated three times and a representative data are presented. *P<0.05, **P<0.01 and ***P<0.001. miR, microRNA; KD, knockdown; OE, overexpression; eEF2K, eukaryotic elongation factor 2 kinase; ATG5, autophagy-related 5; CQ, chloroquine; CT, control; p-, phosphorylated.

Fig 3: The expression of p62 and miR-199a-5p in mice SCLC samples. A–C LC3II/LC3I, ATG5, and p62 expressions in H446 and H446/EP model of SCLC with or without cisplatin treatment. The expression was measured using the western blotting assay. D Representative images of A–C. E Expression of mir-199-5p expressions in H446 and H446/EP model of SCLC with or without cisplatin treatment. F Correlation of the data of all collected tumor samples. (*p < 0.01)

Fig 4: OCSP promotes autophagy but inhibits the expression of ERSR-related proteins(A–F) Representative western blot at the top and densitometric analysis at the bottom of (A) BECN1, (B) ATG5, (C) MAP1LC3B II, (D) SQSTM1, (E) HSPA5, and (F) DDIT3. “L” indicates relatively low PaO2 levels and “H” indicates relatively high PaO2 levels. Data are represented as mean ± SD. p value was calculated using one-way ANOVA. ap < 0.05 versus NMP-L, bp < 0.05 versus NMP-H, cp < 0.05 versus SP-L, dp < 0.05 versus SP-H, ep < 0.05 versus OCSP-L.

Fig 5: Autophagy inhibition in MDR A549/T and H1299/T cells. (A and B) Western blot analysis of LC3I and LC3II protein expression in cells treated with or without 10 µM CQ and the LC3II/LC3I ratio. β-actin was used as the loading control. (C) Neutral red staining for lysosomes analysis. Scale bar, 10 µm. (D) Western blot analysis of the expression levels of PI3K/Akt/mTOR pathway, eEF2K, p62 and ATG5 proteins, and the quantitative analysis results. The experiments were repeated three times, and representative bands are presented. *P<0.05, **P<0.01 and ***P<0.001. eEF2K, eukaryotic elongation factor 2 kinase; ATG5, autophagy-related 5; CQ, chloroquine; CT, control; p-, phosphorylated.