Journal: PeerJ

Article Title: Exosomes from adipose-derived stem cells alleviate premature ovarian failure via blockage of autophagy and AMPK/mTOR pathway

doi: 10.7717/peerj.16517

Figure Lengend Snippet: (A) Ovarian tissue samples were collected on the 15th day, and the apoptosis of ovarian GCs was detected by TUNEL staining. Scale bar: 20 µm. (B) The ultrastructure and intracellular autophagy of cells in ovarian tissue were observed by transmission electron microscope. Scale bar: 2 µm. (C) Levels of autophagy-related genes (Beclin-1 and LC3II/LC3I), apoptosis-related protein Bcl-2, and pathway-related proteins (p-AMPK/AMPK and p-mTOR/mTOR) were detected by western blotting. ** p < 0.01 compared with the WT group. # p < 0.05 and ## p < 0.01 compared with the POF group. Ovarian tissue samples were collected on the 15th day.

Article Snippet: The primary antibodies used in this study were the AMPK antibody (1:1,000, ab32047, Abcam, Cambridge, UK), p-AMPK antibody (1:1,000, ab32047, Abcam), mTOR antibody (1:1,000, CSB-PA208208; Cusabio, Hubei, China), p-mTOR antibody (1:1,000, CSB-PA271384; Cusabio), Beclin-1 antibody (1:1,000, ab210498; Abcam), LC3II/LC3I antibody (1:1,000, 12741T; CST), Bcl-2 antibody (1:1,000, ab182858; Abcam), CD63 antibody (1:1,000, ab217345; Abcam), CD81 antibody (1:1,000, DF2306; Affinity, Cincinnati, OH, USA), TSG101 antibody (1:1,000, DF8427; Affinity), HSP70 antibody (1:1,000, AF5466, Affinity) and the GAPDH antibody (1:1,000, ab245355; Abcam).

Techniques: TUNEL Assay, Staining, Transmission Assay, Microscopy, Western Blot

Journal: PeerJ

Article Title: Exosomes from adipose-derived stem cells alleviate premature ovarian failure via blockage of autophagy and AMPK/mTOR pathway

doi: 10.7717/peerj.16517

Figure Lengend Snippet: (A) ELISA was used to evaluate the levels of E2, FSH, MDA, ROS, and SOD. (B) Ovarian tissue samples were collected on the 15th day, and the apoptosis of ovarian GCs was detected by TUNEL staining. Scale bar: 20 µm. (C–D) The protein levels of Beclin-1, LC3II/LC3I, Bcl-2, p-AMPK, and p-mTOR were detected by western blotting. ** p < 0.01 compared with the WT group, # p < 0.05 and ## p < 0.01 compared with the POF group, and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\hat {}\hat {}$\end{document} ˆ ˆ p < 0.05 and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\hat {}\hat {}$\end{document} ˆ ˆ p < 0.01 compared with the ADSCs group.

Article Snippet: The primary antibodies used in this study were the AMPK antibody (1:1,000, ab32047, Abcam, Cambridge, UK), p-AMPK antibody (1:1,000, ab32047, Abcam), mTOR antibody (1:1,000, CSB-PA208208; Cusabio, Hubei, China), p-mTOR antibody (1:1,000, CSB-PA271384; Cusabio), Beclin-1 antibody (1:1,000, ab210498; Abcam), LC3II/LC3I antibody (1:1,000, 12741T; CST), Bcl-2 antibody (1:1,000, ab182858; Abcam), CD63 antibody (1:1,000, ab217345; Abcam), CD81 antibody (1:1,000, DF2306; Affinity, Cincinnati, OH, USA), TSG101 antibody (1:1,000, DF8427; Affinity), HSP70 antibody (1:1,000, AF5466, Affinity) and the GAPDH antibody (1:1,000, ab245355; Abcam).

Techniques: Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining, Western Blot

Journal: PeerJ

Article Title: Exosomes from adipose-derived stem cells alleviate premature ovarian failure via blockage of autophagy and AMPK/mTOR pathway

doi: 10.7717/peerj.16517

Figure Lengend Snippet: The protein levels of p-AMPK/AMPK, p-mTOR/mTOR, Bcl-2, Beclin-1, and LC3II/LC3I were detected by western blotting. ** p < 0.01 compared with the PBS group, # p < 0.05 and ## p < 0.01 compared with the Exo group.

Article Snippet: The primary antibodies used in this study were the AMPK antibody (1:1,000, ab32047, Abcam, Cambridge, UK), p-AMPK antibody (1:1,000, ab32047, Abcam), mTOR antibody (1:1,000, CSB-PA208208; Cusabio, Hubei, China), p-mTOR antibody (1:1,000, CSB-PA271384; Cusabio), Beclin-1 antibody (1:1,000, ab210498; Abcam), LC3II/LC3I antibody (1:1,000, 12741T; CST), Bcl-2 antibody (1:1,000, ab182858; Abcam), CD63 antibody (1:1,000, ab217345; Abcam), CD81 antibody (1:1,000, DF2306; Affinity, Cincinnati, OH, USA), TSG101 antibody (1:1,000, DF8427; Affinity), HSP70 antibody (1:1,000, AF5466, Affinity) and the GAPDH antibody (1:1,000, ab245355; Abcam).

Techniques: Western Blot

Journal: PeerJ

Article Title: Exosomes from adipose-derived stem cells alleviate premature ovarian failure via blockage of autophagy and AMPK/mTOR pathway

doi: 10.7717/peerj.16517

Figure Lengend Snippet: (A) IC50 value for ADSCs-Exo treating CTX-treated KGN cells was identified by cell counting kit (CCK)-8 assay. (B) Cell viability was detected by CCK-8 assay. (C) Cell apoptosis was assessed by flow cytometry. (D) The protein levels of Beclin-1, LC3II/LC3I, Bcl-2, p-AMPK/AMPK, and p-mTOR/mTOR were detected by western blotting. KGN cells were treated with 250 µM CTX, 10 µg/mL ADSCs-Exo, or/and 5 µM rapamycin (Rapa). ** p < 0.01 compared with the control group, ## p < 0.01 compared with the CTX group, and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\hat {}$\end{document} ˆ p < 0.05 and \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\hat {}\hat {}$\end{document} ˆ ˆ p < 0.01 compared with the CTX + ADSCs-Exo group.

Article Snippet: The primary antibodies used in this study were the AMPK antibody (1:1,000, ab32047, Abcam, Cambridge, UK), p-AMPK antibody (1:1,000, ab32047, Abcam), mTOR antibody (1:1,000, CSB-PA208208; Cusabio, Hubei, China), p-mTOR antibody (1:1,000, CSB-PA271384; Cusabio), Beclin-1 antibody (1:1,000, ab210498; Abcam), LC3II/LC3I antibody (1:1,000, 12741T; CST), Bcl-2 antibody (1:1,000, ab182858; Abcam), CD63 antibody (1:1,000, ab217345; Abcam), CD81 antibody (1:1,000, DF2306; Affinity, Cincinnati, OH, USA), TSG101 antibody (1:1,000, DF8427; Affinity), HSP70 antibody (1:1,000, AF5466, Affinity) and the GAPDH antibody (1:1,000, ab245355; Abcam).

Techniques: Cell Counting, CCK-8 Assay, Flow Cytometry, Western Blot, Control

Journal: Aging (Albany NY)

Article Title: Geniposide-mediated protection against amyloid deposition and behavioral impairment correlates with downregulation of mTOR signaling and enhanced autophagy in a mouse model of Alzheimer's disease

doi: 10.18632/aging.101759

Figure Lengend Snippet: Geniposide treatment decreases mTOR activation markers in brains of APP/PS1 mice. Hippocampal expression of Akt, mTOR, and 4E-BP1, and their respective phosphorylated forms was detected by western blot. The expression of p-Akt ( A ) and p-mTOR ( B ) was enhanced in APP/PS1 mice compared to WT, and geniposide attenuated this increase. The expression of p-4E-BP1 ( C ) in APP/PS1 mice was reduced compared to WT, and geniposide partly restored this decrease. Data are presented as mean ± SEM (n = 6). *** p < 0.001, ** p < 0.01, * p < 0.05 vs. WT; # p < 0.05 vs. APP/PS1 mice (one-way ANOVA, Tukey's Multiple Comparison Test). WT: wild-type mice. GP: geniposide.

Article Snippet: The membranes were blocked in 5% bovine serum albumin in TBST (Tris-buffered saline with 0.05% Tween-20) for 1h, and incubated overnight at 4°C with primary antibodies directed against: Akt (1:1,000), p-Akt (1:2,000), mTOR (1:1,000), p-mTOR (1:1,000), 4E-BP1 (1:1,000), or p-4E-BP1 (1:2,000), followed by incubation at 4°C for 2h with a goat-anti-rabbit IgG-horseradish peroxidase-conjugated secondary antibody (Boster, Wuhan, China).

Techniques: Activation Assay, Expressing, Western Blot, Comparison

Journal: World Journal of Gastroenterology

Article Title: Antagonizing adipose tissue-derived exosome miR-103-hepatocyte phosphatase and tensin homolog pathway alleviates autophagy in non-alcoholic steatohepatitis: A trans-cellular crosstalk

doi: 10.3748/wjg.v29.i29.4528

Figure Lengend Snippet: The capacity of miR-103 in targeting phosphatase and tensin homolog gene and affecting autophagy. A: The luciferase reporter assay results verified the interaction between miR-103 and phosphatase and tensin homolog; B: Western blotting showed the differential expression of the autophagy-related protein; C: Transmission electron microscopy images of autophagosomes (red arrowhead) in the liver; D: The results of immunofluorescence staining showed liver autophagosomes in different groups. a P < 0.01 vs control; b P < 0.01 vs model; c P < 0.01 vs miR-103; d P < 0.05 vs model. PTEN: Phosphatase and tensin homolog; WT: Wild type; MUT: Mutant; DAPI: 4’,6-diamidino-2-phenylindole; mTOR: Mammalian target of rapamycin; NC: Negative control.

Article Snippet: The membranes were blocked and incubated overnight with antibodies against PTEN (9188T, CST), p-AMPK (ab32047, Abcam), p-mammalian target of rapamycin (mTOR) (CSB-PA271384, Cusabio), LC3 (12741T, CST), p62 (ab91526, Abcam), and GAPDH (ab245355, Abcam) at 4 °C.

Techniques: Luciferase, Reporter Assay, Western Blot, Quantitative Proteomics, Transmission Assay, Electron Microscopy, Immunofluorescence, Staining, Control, Mutagenesis, Negative Control

Journal: World Journal of Gastroenterology

Article Title: Antagonizing adipose tissue-derived exosome miR-103-hepatocyte phosphatase and tensin homolog pathway alleviates autophagy in non-alcoholic steatohepatitis: A trans-cellular crosstalk

doi: 10.3748/wjg.v29.i29.4528

Figure Lengend Snippet: The effect of adipose tissue-derived exosomes miR-103 on autophagy in mice. A and B: Western blotting detected the expression of the autophagy-related protein; C: The results of immunofluorescence staining to observe autophagosomes in the liver from different groups; D: Transmission electron microscopy images of autophagosomes (red arrowhead) in the liver from different groups. a P < 0.05 vs control; b P < 0.01 vs control; c P < 0.05 vs exosomes; d P < 0.01 vs exosomes. Exo: Exosomes; PTEN: Phosphatase and tensin homolog; DAPI: 4’,6-diamidino-2-phenylindole; mTOR: Mammalian target of rapamycin; NC: Negative control.

Article Snippet: The membranes were blocked and incubated overnight with antibodies against PTEN (9188T, CST), p-AMPK (ab32047, Abcam), p-mammalian target of rapamycin (mTOR) (CSB-PA271384, Cusabio), LC3 (12741T, CST), p62 (ab91526, Abcam), and GAPDH (ab245355, Abcam) at 4 °C.

Techniques: Derivative Assay, Western Blot, Expressing, Immunofluorescence, Staining, Transmission Assay, Electron Microscopy, Control, Negative Control