lc3 ii  (Bioss)

Journal: Redox Biology

Article Title: YY1 nitration participates in DbCM cardiomyocyte lipotoxicity by inhibiting ANXA3 -induced microlipophagy

doi: 10.1016/j.redox.2026.104085

Figure Lengend Snippet: ANXA3 did not regulate myocardial free fatty acid uptake or lipolysis but positively modulated macroautophagy in T2DM mice. (A-C) Relative p62 protein expression level and LC3 II/I ratio in heart tissues of db/m and db/db mice analyzed by Western blot, n = 4. (D-F) Relative p62 protein expression level and LC3 II/I ratio in heart tissues of mice analyzed by Western blot, n = 4. (G-I) Relative p62 protein expression level and LC3 II/I ratio in AC16 cardiomyocytes analyzed by Western blot, n = 6. (J-L) Relative p62 protein expression level and LC3 II/I ratio in AC16 cardiomyocytes analyzed by Western blot, n = 6-8. The data were presented as the mean ± SD. ∗ P < 0.05 versus the db/m group or the db/db + OE-GFP group; ∗∗ P < 0.01 versus the db/m group or the vehicle group or the OE-NC + vehicle group; ∗∗∗ P < 0.001 versus the vehicle group or the OE-NC + vehicle group; # P < 0.05 versus the OE-NC + HGPA group; ### P < 0.001 versus the OE-NC + HGPA group.

Article Snippet: Blocked with 5% (w/v) non-fat-dried milkat room temperature for 1 h. Then the membranes were incubated with the anti-ANXA3 antibody (Proteintech, 11804-1-AP; 1:1000 [v/v]), the anti-YY1 antibody (Proteintech, 22156-1-AP; 1:1000 [v/v]) the anti-PLIN2 antibody (Proteintech, 15294-1-AP; 1:1000 [v/v]), the anti-SQSTM1/p62 antibody (Cell Signaling Technology, 23214; 1:1000 [v/v]), the anti-LC3 antibody (Cell Signaling Technology, 12741; 1:1000 [v/v]), the anti-Rab7 antibody (Proteintech, 55469-1-AP; 1:1000 [v/v])or the rabbit anti-α-Tubulin antibody (ABclonal, AC031; 1:1000 [v/v])overnight at 4 °C.

Techniques: Expressing, Western Blot

Journal: Genes & Diseases

Article Title: TIGAR promotes osteogenic differentiation and ameliorates glucocorticoid-induced osteoporosis via autophagy-Nrf2-ROS axis

doi: 10.1016/j.gendis.2025.101735

Figure Lengend Snippet: TIGAR activated nuclear factor erythroid-2 related factor (Nrf2) to reduce dexamethasone (Dex)-induced oxidative stress through inducing autophagy. Bone marrow mesenchymal stem cells (BMSCs) were transfected with TIGAR overexpression plasmid and treated with Dex. (A, B) Immunofluorescence staining of Nrf2 of BMSCs and the quantification of the integrated optical density (IOD) per field. Scale bars, 50 μm. (C, D) Western blot analysis and quantification of Nrf2 expression in extracted nuclear proteins. (E, F) ROS level under Dex treatment in BMSCs with or without administration of 10 nM Nrf2 inhibitor (ML385) after transfecting with TIGAR overexpression plasmid, and the quantification of IOD per filed. Scale bars, 100 μm. BMSCs were treated with Dex and chloroquine (CQ) (20 μM) after transfecting with TIGAR overexpression plasmid. (G–I) Western blot analysis and quantification of p62 and LC3-II expression under different treatments. (J, K) Representative images of mCherry-GFP-LC3 puncta and number of autophagosomes (yellow) (analyzed by Pearson's correlation). Scale bars, 50 μm. (L–N) Western blot analysis and quantification of Nrf2 and kelch-associated protein 1 (Keap1) expression under different treatments. (O, P) The immunofluorescence staining of Nrf2 in BMSCs and the quantification of the IOD per field. Scale bars, 50 μm. (Q, R) Representative immunofluorescence images of LC3 and Keap1. Pearson's correlation of co-localization is shown in the bar graph format from the three independent experiments analyzed. Scale bars, 50 μm. (S, T) Representative images of ROS and the quantification of the IOD per field. Scale bars, 100 μm. Data are shown as mean ± SEM. n = 3, biologically independent samples. Two-way analysis of variance (ANOVA) with Tukey's multiple comparisons test was used to assess statistical significance. ∗ p < 0.05, ∗∗ p < 0.01. NC, negative control. OE, TIGAR overexpression plasmid.

Article Snippet: Primary antibodies LC3 (1:100, Cell Signaling Technology, #12741), Keap1(1:100, Zen-bio, R26935 ) and Nrf2 (1:100, Zen-Bio, 380773) were incubated overnight at 4 °C.

Techniques: Transfection, Over Expression, Plasmid Preparation, Immunofluorescence, Staining, Western Blot, Expressing, Negative Control