Fig 2: Andro upregulated the expression of p62 and LC3 proteins and mRNA in PC12 cells induced by Al(mal)3. Cells were incubated with 700 µM Al(mal)3 and 5 or 10 µM Andro for 24 h. The expression of p62 and LC3 proteins were detected by western blot, GAPDH was used as loading control (A). Quantitative analysis of p62 (B) and the ratio of LC3-II to LC3-I (C) protein expression levels. The levels of p62 and LC3 (D) mRNA were analysed using RT-qPCR *p < 0.05, **p < 0.01 versus the control, #p < 0.05, ##p < 0.01 versus Al(mal)3 group was considered statistically significant differences (n = 3).

Fig 3: Effects of SP on the expression levels of Bax, Bcl-2, Beclin-1 and LC3 in heart failure rats. The expression levels of Bcl-2, Beclin-1 and LC3 were reduced in the heart-failure model. SP promoted the expression levels of Bax and LC3, and inhibited Beclin-1 expression. *P<0.05 vs. control group; #P<0.05 vs. model group (n=6 repeats/group). SP, substance P; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; LC3, microtubule-associated protein 1A/1B-light chain 3.

Fig 4: Effects of SP on the expression levels of Bax, Bcl-2, Beclin-1 and LC3 in H9c2 cells. Bcl-2 expression in the doxorubicin-treated group was significantly lower than in the control group, whereas SP did not influence Bcl-2 expression in the doxorubicin-treated group. Doxorubicin reduced the expression levels of Beclin-1 and LC3 in H9c2 cells, whereas SP further reduced Beclin-1 expression, but inhibited the doxorubicin-induced decrease in LC3 levels. *P<0.05 vs. control cells; #P<0.05 vs. model cells (n=6 repeats in each group). SP, substance P; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; LC3, microtubule-associated protein 1A/1B-light chain 3.

Fig 5: Metformin treatment upregulates the expression of LC3 in the liver of HFD-induced obese mice. (A) Microphotographs of liver tissues from mice in the normal chow diet, high fat diet and high fat diet+metformin groups following immunohistochemical staining of LC3. (B) Quantitative analysis of LC3-positive regions in the liver tissue sections. (C) mRNA levels of LC3 in the liver, measured using reverse transcription-quantitative polymerase chain reaction analysis and normalized to glyceraldehyde-3-phosphate dehydrogenase. Data are presented as the mean ± standard error of the mean **P<0.01; ***P<0.001. LC3, microtubule-associated protein 1 light chain 3.