Journal: Biochemistry and Biophysics Reports

Article Title: Screening of metabolic-related biomarkers linking intervertebral disc degeneration and type 2 diabetes based on comprehensive bioinformatics analysis and machine learning

doi: 10.1016/j.bbrep.2026.102593

Figure Lengend Snippet: BCAA metabolism and immune changes in degenerated NP cells. (A) qRT-PCR was conducted to analysis the mRNA levels of BCAA metabolism-related enzymes and BCAA transporter in grade I/II and grade III/IV NP tissues; n = 20; P < 0.05. (B, C) Expression levels of mRNA for pro-inflammatory factors, BCAA metabolism-related enzymes, and BCAA transporters in control group and TNF-α-treated HNPC cells. (D) Expression of SA-β-gal in HNPC cells stimulated with different concentrations of TNF-α. (E, F) p16, p21, p53 mRNA level in TNF-α-stimulated HNPC cells,and correlation analysis with TNF-α concentration.

Article Snippet: Cellular senescence was assessed using Senescence-Associated β-Galactosidase (SA-β-gal) staining (Senescence β-Galactosidase Staining Kit, #C0602, Beyotime, China), performed according to the manufacturer's protocol.

Techniques: Quantitative RT-PCR, Expressing, Control, Concentration Assay

Journal: Redox Biology

Article Title: FTMT-mediated suppression of mitophagy links iron accumulation to osteoporosis

doi: 10.1016/j.redox.2026.104157

Figure Lengend Snippet: Iron accumulation impairs mitophagy, promotes senescence, and suppresses osteogenic differentiation in BMSCs. (a) Schematic diagram of extraction of BMSCs from human femur. (b) Western blot analysis of osteogenic marker proteins (RUNX2, ALP) in BMSCs from normal controls and postmenopausal osteoporosis patients and osteoporosis patients with iron accumulation. (c) Alizarin Red S (ARS) staining of BMSCs treated with increasing concentrations of FAC (0, 50, 100, 200 μM) for 21 days and alkaline phosphatase (ALP) staining of BMSCs treated with increasing concentrations of FAC (0, 50, 100, 200 μM) for 14 days. Scale bar: 50 μm. (d) Western blot analysis of osteogenic markers (RUNX2, ALP) in FAC-treated BMSCs for 5 days. (e) RT-qPCR analysis of osteogenic genes ( Runx2, Alpl, Bglap, Sp7 ) in FAC-treated BMSCs for 72h. (f) KEGG pathway enrichment analysis of differentially expressed genes from RNA sequencing of control and 200 μM FAC-treated BMSCs for 72h. (g, h) Immunofluorescence staining of senescence markers (γ-H2AX, H3K9me3) in FAC-treated BMSCs for 72h. Scale bar: 20 μm. (i) Senescence-associated β-galactosidase (SA-β-gal) staining of FAC-treated BMSCs for 72h. Scale bar: 50 μm. (j) Flow cytometric quantification of SA-β-gal activity in FAC-treated BMSCs for 72h. (k) Western blot analysis of senescence-related proteins (P53, P21, P16) in FAC-treated BMSCs for 72h. (l) Mitophagy assessment by immunofluorescence co-staining with Mitophagy Dye (red) and MitoTracker (green) in FAC-treated BMSCs for 72h. Scale bar: 20 μm. (m) Western blot analysis of mitophagy/autophagy-related proteins (PINK1, PARKIN, P62, LC3) in FAC-treated BMSCs for 72h. (n) Mitochondrial membrane potential (MMP) detection by MT-1 staining in FAC-treated BMSCs for 72h. Scale bar: 30 μm. Data are presented as mean ± SEM; One-way ANOVA (Dunnett's multiple-comparison test); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: After treatment for 72 h, the BMSCs were fixed with 4% formaldehyde and stained with a Senescence β-Galactosidase Staining Kit (BeyoTime, C0602) following the protocol published by the manufacturer. β-Galactosidase quantification.

Techniques: Extraction, Western Blot, Marker, Staining, Quantitative RT-PCR, RNA Sequencing, Control, Immunofluorescence, Activity Assay, Membrane, Comparison