Journal: Journal of Ovarian Research

Article Title: MSCs–derived EVs protect against chemotherapy-induced ovarian toxicity: role of PI3K/AKT/mTOR axis

doi: 10.1186/s13048-024-01545-7

Figure Lengend Snippet: Effect of MSCs-EVs, rapamycin, and quercetin on ( A ) phosphorylation of mTOR, PI3K, and AKT as detected by western blot analysis, ( B - D ) Intensity of immunoreactivity of selected proteins as quantified by densitometry. Results are expressed as mean ± SEM. **significant vs. Control group at p<0.01, **** at p<0.0001, # significant vs. OF group at p<0.05, ## at p<0.01, ### at p<0.001, #### at p<0.0001, & significant vs. OF + Rapamycin group at p<0.05

Article Snippet: Next, the blots were incubated with the appropriate primary antibodies for an entire night at 4 °C, PTEN (E-AB-63495, Elabscience, USA), FOXO3 (NBP2-16521, Novus Biologicals USA), mTOR (sc-517464, Santa Cruz Biotechnology, USA), Phospho-mTOR (sc-293133, Santa Cruz Biotechnology, USA), PI3K (E-AB-64202, Elabscience, USA), Phospho-PI3K (E-AB-20966, Elabscience, USA), AKT (E-AB-15441, Elabscience, USA), Phospho-AKT (E-AB-20804, Elabscience, USA), β-actin (E-AB-20031, Elabscience, USA).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of hepatology

Article Title: Hepatic stellate cell activation promotes alcohol-induced steatohepatitis through Igfbp3 and SerpinA12

doi: 10.1016/j.jhep.2020.02.005

Figure Lengend Snippet: (A) Representative images and BODIPY® staining quantification show pretreatment with SerpinA12 protects against lipid droplet formation induced by EtOH (n=6, 63x, p<0.05, one way ANOVA). (B) WB and densitometry quantification shows that EtOH induces a reduction in the phosphorylation of AMPK compared to control. SerpinA12 induces phosphorylation of AMPK, even in the presence of EtOH, and this effect is blunted by the presence of AMPK inhibitor (duplicates, n=6, *p<0.05, one way ANOVA). (C) Representative images and BODIPY staining quantification show protection of SerpinA12 against EtOH-induced lipid droplet formation on treatment with SerpinA12 ± EtOH. This effect was blunted by using an AMPK inhibitor (n=6, 63x, *p<0.05, one way ANOVA). (D) EtOH induces key lipogenic genes involved in hepatic lipid homeostasis, and this effect is reduced by SerpinA12. The protective effect of SerpinA12 is attenuated by an AMPK inhibitor (n=6, *p<0.05 compared to control, #p<0.05 compared to EtOH alone, one way ANOVA). (E) Human hepatocytes treated with supernatant from human HSC with a siRNA-mediated knockdown of SerpinA12 ± EtOH. EtOH ± supernatant from siRNA-mediated knockdown of SerpinA12 increases expression of SREBP-1c, FASN, and SCD1 compared to EtOH in siControl cells (n=9, *p<0.05 compared to control, #p<0.05 compared to EtOH alone, one way ANOVA).

Article Snippet: Compounds used were murine or human recombinant Igfbp3 (R&D systems, 0.3 μg/mL), Cytochalasin D (Sigma-Aldrich, 1:1000), PP2 (Sigma-Aldrich, 10 uM), recombinant SerpinA12 (R&Dsystems, 100 ng/mL), Akt inhibitor (Tocris, Akti-1/2, 250 nM), PQ-401 (Tocris, 10uM) and AMPK inhibitor (Tocris, Dorsomorphin dihydrochloride, 20 uM).

Techniques: Staining, Expressing

Journal: Journal of hepatology

Article Title: Hepatic stellate cell activation promotes alcohol-induced steatohepatitis through Igfbp3 and SerpinA12

doi: 10.1016/j.jhep.2020.02.005

Figure Lengend Snippet: (A) Representative BODIPY® staining images and quantification from HepG2Cyp2E1 cells treated with (I) medium, (II) control HSC supernatant, (III) siRNA mediated NRP-1 knockdown HSC supernatant. (III) shows decreased lipid droplet formation in vitro (n=6–9, 63X, one-way ANOVA). (B) Top ten down/upregulated molecules from adipokine and inflammatory protein array from NRP-1 KD HSC supernatant compared to controls. IGFBP3 and SerpinA12 were the most significantly decreased and increased proteins respectively (n=3, p<0.05, unpaired t-test). (C) Igfbp3 mRNA and protein expression in NRP-1 KD HSC is lower than human WT HSC on qPCR and WB (n=6–9, p<0.05, unpaired t-test). (D) SerpinA12 mRNA expression in NRP-1 KD HSC is higher than human WT HSC (n=6–9, p<0.05, unpaired t-test).

Article Snippet: Compounds used were murine or human recombinant Igfbp3 (R&D systems, 0.3 μg/mL), Cytochalasin D (Sigma-Aldrich, 1:1000), PP2 (Sigma-Aldrich, 10 uM), recombinant SerpinA12 (R&Dsystems, 100 ng/mL), Akt inhibitor (Tocris, Akti-1/2, 250 nM), PQ-401 (Tocris, 10uM) and AMPK inhibitor (Tocris, Dorsomorphin dihydrochloride, 20 uM).

Techniques: Staining, In Vitro, Protein Array, Expressing

Journal: Journal of hepatology

Article Title: Hepatic stellate cell activation promotes alcohol-induced steatohepatitis through Igfbp3 and SerpinA12

doi: 10.1016/j.jhep.2020.02.005

Figure Lengend Snippet: (A) Representative WB and densitometry quantification for Igfbp3 and SerpinA12 from plasma from control subjects and with alcoholic hepatitis. Subjects with alcoholic hepatitis have significative higher levels of Igfbp3 and significative lower levels of SerpinA12 compared to controls (n=9, p<0.05, unpaired t-test). (B) Schematic representation of the proposed mechanism of hepatic stellate cells (HSC)-activation-induced steatosis. Activated HSC increase Insulin-like growth factor-binding protein 3 (Igfbp3) and reduces SerpinA12 secretion, which have a paracrine effect over hepatocytes. Igfbp3 increases p-Akt signaling through integrin receptor leading to lipid droplets formation, triglyceride content, and lipogenic gene expression; EtOH reduces p-AMPK, increasing lipogenesis. SerpinA12 protects against ethanol-induced steatosis through increasing p-AMPK.

Article Snippet: Compounds used were murine or human recombinant Igfbp3 (R&D systems, 0.3 μg/mL), Cytochalasin D (Sigma-Aldrich, 1:1000), PP2 (Sigma-Aldrich, 10 uM), recombinant SerpinA12 (R&Dsystems, 100 ng/mL), Akt inhibitor (Tocris, Akti-1/2, 250 nM), PQ-401 (Tocris, 10uM) and AMPK inhibitor (Tocris, Dorsomorphin dihydrochloride, 20 uM).

Techniques: Activation Assay, Binding Assay, Expressing