Journal: International Journal of Molecular Sciences

Article Title: Liraglutide Attenuates FFA-Induced Retinal Pigment Epithelium Dysfunction via AMPK Activation and Lipid Homeostasis Regulation in ARPE-19 Cells

doi: 10.3390/ijms26083704

Figure Lengend Snippet: Figure 3. Liraglutide counteracts FFA-induced autophagy suppression and enhances lipophagy in ARPE-19 cells. (A) AO staining of ARPE-19 cells treated with FFA (250 µM) and/or liraglutide (0.1 µM) for 24 h. FFA significantly reduced the expression of AVOs, indicating suppressed autophagy, while liraglutide co-treatment restored AVO expression. Scale bar = 50 µm. (B) HCA quantification of AVOs. FFA reduced the number of AVOs to ~10% of control levels, while liraglutide co-treatment recovered AVOs to ~50% of control levels. (C) Western blot analysis of pThr172-AMPK, LC3-II, p62, and pSer2448- mTOR in ARPE-19 cells. FFA significantly decreased pThr172-AMPK and LC3-II expression and increased p62 levels, consistent with autophagy suppression. Additionally, FFA markedly elevated p-mTOR levels, indicating mTOR activation and the further inhibition of autophagy. Liraglutide co-treatment reversed these effects, restoring pThr172-AMPK and LC3-II levels while reducing p62 accumulation, and suppressing mTOR phosphorylation, suggesting the activation of AMPK-mediated autophagy through inhibition of the mTOR pathway. (D) Immunocytochemistry staining of PLIN2 in ARPE-19 cells. FFA markedly reduced PLIN2 expression, while liraglutide restored it. Co-localization of PLIN2 with LDs suggests that liraglutide promotes lipophagy and enhances LD degradation. Scale bar = 20 µm. (E) Quantitative PCR analysis of lipophagy and lysosomal degradation-related genes, including autophagy related 5 (ATG5), lysosomal acid lipase (LIPA), ras-related protein 7A (RAB7A), and lysosomal-associated membrane protein 2 (LAMP2). FFA significantly downregulated these genes, indicating suppressed LD degradation pathways, whereas liraglutide restored their expression levels. All data were collected from at least three independent experiments and are presented as mean ± SD. For the HCA quantification of AVOs, a minimum of 100 cells per condition

Article Snippet: Primary antibodies against AMPK, phosphorylated Thr172-AMPK (pThr172AMPK), mTOR, and phosphorylated Ser2448-mTOR (pSer2448-mTOR) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Staining, Expressing, Control, Western Blot, Activation Assay, Inhibition, Phospho-proteomics, Immunocytochemistry, Real-time Polymerase Chain Reaction, Membrane

Journal: Frontiers in Physiology

Article Title: Cardiac implications of chicken wooden breast myopathy

doi: 10.3389/fphys.2025.1547661

Figure Lengend Snippet: Primary antibodies used for immunoblotting.

Article Snippet: pSer2448-mTOR , F1NUX4 F1NUX4_CHICK (2521aa) , 260 kDa , 5% BSA , #2971 , 1:1,000 , Cell Signaling.

Techniques: Western Blot, Blocking Assay

Journal: Current Issues in Molecular Biology

Article Title: Inhibition of eNOS Partially Blunts the Beneficial Effects of Nebivolol on Angiotensin II-Induced Signaling in H9c2 Cardiomyoblasts

doi: 10.3390/cimb44050144

Figure Lengend Snippet: Effects of Neb plus L-N combination on Ang II-induced mTOR signaling in H9c2 cardiomyoblasts. Top: Representative images of autoradiograms showing elevated levels of pSer2448 mTOR ( A ), pThr389 S6K1 ( B ), and pSer235/236RPS6 ( C ) in H9c2 cardiomyoblasts stimulated with Ang II and suppression by Neb and Neb + L-N. Bottom: Graphs show results of densitometry analysis of the intensity of the phosphorylated protein bands after adjusting for the intensity of total protein bands (tmTOR, tS6K1, tRPS6). Top: Representative images of autoradiograms showing elevated levels of pSer2448 mTOR ( D ), pThr389 S6K1 ( E ), and pSer235/236RPS6 ( F ) in H9c2 cardiomyoblasts stimulated with Ang II and suppression by Rap. Bottom: Graphs show results of densitometry analysis of the intensity of the phosphorylated protein bands after adjusting for the intensity of total protein bands (tmTOR, tS6K1, tRPS6). * p < 0.05 vs. untreated (Con) and # p < 0.05 vs. Ang II. Values are means ± SEM. N ≥ 3–6 for each treatment group.

Article Snippet: Antibodies for phospho-mTOR (pSer2448), mTOR, phospho-p70S6K (pThr389), p70S6K, phospho-RPS6 (pSer235/236), and RPS6 were obtained from Cell Signaling Technology Inc. (Boston, MA, USA).

Techniques: