Journal: Journal of Translational Medicine

Article Title: Targeting the PINK1/Parkin-FNDC5 pathway: a novel mechanism of icariin in regulating muscle-bone metabolic coupling in osteosarcopenia

doi: 10.1186/s12967-026-08017-0

Figure Lengend Snippet: A : JC-1 reagent detection results; B : JC-1 monomer ratio; C : PINK1 protein expression histogram; D : Parkin protein expression histogram; E : LC3 protein expression histogram; F : p62 protein expression histogram; G : Protein bands; H : PINK1 mRNA expression; I : Parkin mRNA expression; J : LC3 mRNA expression; K : p62 mRNA expression; Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. Data are expressed as mean ± SD. ns (not significant), * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: After closing, incubate the membrane with anti-PINK1 (1:1000, Bioss, BS-22173R), Parkin (1:1000, Bioss, BS-23687R), LC3 (1:500, Solarbio, K008014P), p62 (1:1000, Bioss, BS-55207R), FNDC5 (1:1000, Bioss, BS-8486R), Runx2 (1:1000, BS-1134R), OCN (1:500, BS-4917R), Actin (1:20,000, Bioss, BS-10966R) as the internal control.

Techniques: Expressing

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: These are the primary antibodies that were used in this study: anti-mouse TIGAR (1:500, Santa Cruz, sc166290), anti-rabbit Runx2 (1:1000, Cell Signaling Technology, #12556), anti-rabbit sp7 (1:1000, Abcam, ab209484), anti-rabbit Nrf2 (1:1000, Zen-Bio, 380773), anti-rabbit keap1 (1:1000, Zen-Bio, R26935 ), anti-rabbit p62 (1:1000, Proteintech, 18420-1-AP), anti-rabbit LC3A/B (1:1000, Cell Signaling Technology, #12741), anti-rabbit HDAC1(1:1000, Cell Signaling Technology, #34589), and anti-mouse GAPDH (1:10000, ABclonal, AC002).

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