Journal: Molecular Neurobiology

Article Title: Comparative Analysis of Red Onion-Derived Exosome-Like Nanovesicles and Extract Reveals Sustained Immunomodulatory Effects in LPS/IFN-γ-Stimulated Microglia

doi: 10.1007/s12035-026-05820-0

Figure Lengend Snippet: Anti-inflammatory and antioxidant properties of RO-ELNs and extract against ATP + LPS/IFN-γ stimulated pro-inflammatory mediators in HMC3 cells. RO-ELNs with a concentration of 1 × 10 9 particles/mL were pretreated for 20 or 6 h before acute or prolonged neuroinflammation models. Extract with a concentration of 125 µg/mL was incubated for only 6 h for both acute and prolonged neuroinflammation models. Western blot and quantitative analyses show the ability of RO-ELNs and extract in inhibiting P-NF-κB ( A , B ) and COX-2 ( C , D ) expression levels in HMC3 cells at different time points 4 and 48 h of neuroinflammation. GAPDH was used as a housekeeping protein. The antioxidative properties of RO-ELNs and extract ( E ) and their ability to decrease IL-6, TNF-α secretion levels ( F , G ) at 4 and 48 h of neuroinflammation. Data are expressed as mean ± standard ( n ≥ 3): * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. As a result of the initial reversed loading sequence for COX-2, lanes were rearranged digitally to align with the experimental grouping presented in the paper. All lanes originated from the same membrane and exposure. A separation dash marks the realignment. Uncropped original membranes can be found in Supplementary Figure

Article Snippet: 3033), and GAPDH (Boster, M00227-7) antibodies overnight at 4 °C.

Techniques: Concentration Assay, Incubation, Western Blot, Expressing, Sequencing, Membrane

Journal: iScience

Article Title: Role of Fyn and the interleukin-6-STAT-3-autophagy axis in sarcopenia

doi: 10.1016/j.isci.2023.107717

Figure Lengend Snippet: Muscular autophagy in the tail-suspension secondary sarcopenia model (A) Selective muscle sizes from control and TS mice (n = 6 per group). Data are expressed as mean ± SEM. ∗∗∗p < 0.005 vs. control. (B) mRNA expression levels of atrogin-1 and MuRF-1, relative to β-actin, in the gastrocnemius muscles of TS mice (n = 3). Data are expressed as mean ± SEM. ∗∗∗p < 0.005 vs. control. (C) Representative immunoblots from three independent experiments with gastrocnemius muscles of control and TS mice, developed using the indicated antibodies. Signal quantification of the protein expression levels of Fyn, normalized to GAPDH (n = 3). Data are shown as mean ± SEM. ∗∗∗p < 0.005 vs. control. Signal quantification of the expression levels of total STAT3, normalized to GAPDH, and phosphoY705-STAT3, normalized with total STAT3 (n = 3). Data are shown as mean ± SEM. ∗∗∗p < 0.005 vs. control. (D) Representative H&E staining of control and TS mice. See also Figure S2 . (E) Representative p62 immunofluorescence visualized in control and TS mice. Proportion of p62-positive myocytes in control and TS mice (n = 3). Data are expressed as mean ± SEM. ∗p < 0.05 vs. control. (F) Three-month-old control and TS mice were treated with either vehicle (as control) or 0.4 mg/kg/day colchicine for two days. Immunoblots of the TA muscle were obtained using the indicated antibodies. Representative immunoblots of three independent experiments. Signal quantification of the expression levels of LC3-II, normalized with GAPDH levels (n = 3). Data are expressed as mean ± SEM. ∗∗∗p < 0.005 vs. control. The Mann–Whitney U test was used for statistical comparison. The bars on each column show standard error of the mean. Scale bars in the tissue images represent the lengths defined in each figure. Abbreviations: TA, tibialis anterior; TS, tail suspension; SEM, standard error of the mean; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; STAT3, signal transducer and activator of transcription 3; C, control; Col, colchicine.

Article Snippet: Anti-GAPDH mouse monoclonal antibody was obtained from MBL Co., Ltd. (Nagoya, Japan).

Techniques: Suspension, Control, Expressing, Muscles, Western Blot, Staining, Immunofluorescence, MANN-WHITNEY, Comparison

Journal: iScience

Article Title: Role of Fyn and the interleukin-6-STAT-3-autophagy axis in sarcopenia

doi: 10.1016/j.isci.2023.107717

Figure Lengend Snippet: Fyn-dependent STAT3 regulation of autophagy (A) Representative immunoblots from three independent experiments. After 15 min of IL-6 treatment, lysates of C2C12 myotubes were immunoprecipitated with Fyn antibody and immunoblots were prepared using the indicated antibodies. (B) Representative immunoblots from three independent experiments. After 15 min of either IL-1β or IL-6 treatment, lysates of C2C12 myotubes were immunoprecipitated with total STAT3 antibody, followed by immunoblotting with Fyn. (C) Representative immunoblots from three independent experiments indicating autophagy flux in myotube lysates, determined after NH 4 Cl/leupeptin treatment for 2 h, followed by IL-6 treatment. Lysates were immunoblotted with the indicated antibodies (a). Signal quantification of the expression levels for LC3-II, normalized to GAPDH (n = 3) (b). Data are expressed as mean ± SEM. ∗p < 0.05, ∗∗∗p < 0.005 vs. control. The Mann-Whitney U test was used for statistical comparison. The bars on each column show standard error of the mean. Abbreviations: STAT3, signal transducer and activator of transcription; IL, interleukin; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; SEM, standard error of the mean; N/L, NH 4 Cl/leupeptin.

Article Snippet: Anti-GAPDH mouse monoclonal antibody was obtained from MBL Co., Ltd. (Nagoya, Japan).

Techniques: Western Blot, Immunoprecipitation, Expressing, Control, MANN-WHITNEY, Comparison

Journal: iScience

Article Title: Role of Fyn and the interleukin-6-STAT-3-autophagy axis in sarcopenia

doi: 10.1016/j.isci.2023.107717

Figure Lengend Snippet: Fyn-dependent autophagy in muscles with sarcopenia (A) Representative H&E staining of WT and Fyn-KO mice (TS and control). (B) The size distribution of myofibers of WT (upper panel) and Fyn-KO (lower panel) mice with TS (black bar) or control (white bar). (C) Representative p62 immunofluorescence visualized in WT and Fyn-KO mice (TS and control). Proportion of p62-positive myocytes of WT and Fyn-KO mice (TS and control; n = 3). Data are expressed as mean ± SEM. ∗p < 0.05 vs. control. (D) Representative immunoblots from three independent experiments where lysates of three-month-old control and TS mice were treated with either vehicle or 0.4 mg/kg/day colchicine for two days and TA tissues were immunoblotted with the indicated antibodies. Signal quantification of the expression levels of LC3-II, normalized to GAPDH (n = 3). Data are expressed as mean ± SEM. ∗p < 0.05 vs. control. The Mann–Whitney U test was used for statistical comparison between groups. The bars on each column show standard error of the mean. Scale bars in the tissue images represent the lengths defined in each figure. Abbreviations: H&E, hematoxylin & eosin; WT, wild-type; Fyn-KO, Fyn knockout; TS, tail suspension; SEM, standard error of the mean; TA, tibialis anterior; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. C, control, Col, colchicine.

Article Snippet: Anti-GAPDH mouse monoclonal antibody was obtained from MBL Co., Ltd. (Nagoya, Japan).

Techniques: Muscles, Staining, Control, Immunofluorescence, Western Blot, Expressing, MANN-WHITNEY, Comparison, Knock-Out, Suspension