Journal: bioRxiv

Article Title: XL-MS and De Novo Protein Design Identified a Common Motif for TREM2 Binding

doi: 10.64898/2026.04.23.720433

Figure Lengend Snippet: (a) SDS-PAGE analysis showing successful cross-linking of the heterodimeric TREM2 ECD /Trx-ApoE3 complex. (b) Representative high-quality MS/MS spectra of inter-protein cross-linked peptides. (c) Bar representation showing intra-protein XLs, inter-protein XLs, and inter-protein self-links. Figure was created using xiNET . ApoE3: light blue (N-terminal region, residues 1-167), light yellow (hinge region, residues 168-205), and pink (C-terminal region, residues 206-299).

Article Snippet: 20 μM Trx-ApoE3 (Sino Biological 10817-H30E) and 20 μM TREM2 ectodomain were incubated for 1 h at 4°C.

Techniques: SDS Page, Tandem Mass Spectroscopy

Journal: European Journal of Medical Research

Article Title: High glucose induces osteoblasts ferroptosis via the TXNIP/Trx-1/GPX4 pathway in type 2 diabetic osteoporosis

doi: 10.1186/s40001-025-03749-z

Figure Lengend Snippet: TXNIP silencing attenuates ferroptosis and bone loss in T2DOP rats. Forty male SD rats were randomly allocated into four groups (Ctrl, Ctrl + si-TXNIP, T2DOP, and T2DOP + si-TXNIP; n = 10/group). A A schematic illustrates the T2DOP induction protocol, including high-fat diet feeding, STZ administration, and tail vein delivery of TXNIP-siRNA. B TXNIP protein expression in femoral tissue was assessed by Western blot. C, D T2DOP rats exhibited elevated serum iron and MDA levels, both of which were reduced by TXNIP silencing. E Serum GSH levels were significantly decreased in T2DOP rats and partially restored following si-TXNIP treatment. F , G Serum ALP and P1NP levels were measured as indicators of osteogenic activity. H , I Micro-CT analysis revealed severe deterioration of femoral trabecular microarchitecture in T2DOP rats, with improvements in BMD, Tb.N, Tb.Th, and BV/TV following TXNIP knockdown. J Western blot analysis showed downregulation of Trx-1 and GPX4 in T2DOP rats and restoration of both proteins after TXNIP silencing. K Immunohistochemical staining of TXNIP, Trx-1, and GPX4 in femoral sections (scale bar = 100 μm), and L corresponding quantitative analyses further confirmed elevated TXNIP expression and reduced Trx-1/GPX4 levels in T2DOP rats, with these changes reversed by si-TXNIP. Data are presented as mean ± SD. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Equal amounts of protein were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, IPVH00010), blocked with 5% BSA, and incubated overnight at 4 °C with primary antibodies against TXNIP (ab188865, Abcam, RRID:AB_2923036), Trx-1 (A4024, ABclonal, RRID:AB_2863174), GPX4 (ab125066, Abcam, RRID:AB_10973901), and β-actin (ab115777, Abcam, RRID:AB_10899528).

Techniques: Expressing, Western Blot, Activity Assay, Micro-CT, Knockdown, Immunohistochemical staining, Staining

Journal: European Journal of Medical Research

Article Title: High glucose induces osteoblasts ferroptosis via the TXNIP/Trx-1/GPX4 pathway in type 2 diabetic osteoporosis

doi: 10.1186/s40001-025-03749-z

Figure Lengend Snippet: High glucose upregulates TXNIP and promotes ferroptosis by suppressing Trx-1–mediated antioxidant defense in osteoblasts. A Heatmap illustrating global transcriptional alterations in hFOB1.19 cells following NG or HG treatment. B Volcano plot identifying TXNIP as one of the most significantly upregulated genes under HG exposure. C KEGG pathway enrichment analysis revealing marked activation of ferroptosis-associated signaling pathways in the HG group. D , E qRT-PCR and Western blot analyses demonstrating a robust increase in TXNIP mRNA and protein expression in HG-treated cells. F , G HG exposure significantly decreased Trx-1 expression at both the mRNA and protein levels. H Co-immunoprecipitation confirming enhanced TXNIP–Trx-1 interaction under HG conditions. I Western blot verifying efficient TXNIP silencing in hFOB1.19 cells. J TXNIP knockdown restored Trx-1 and GPX4 expression in HG-treated osteoblasts. K Western blot showing that TXNIP silencing reversed the HG-induced suppression of TrxR1 protein. L Measurement of TrxR1 activity demonstrating reduced enzymatic activity under HG conditions and partial recovery upon TXNIP knockdown. M Western blot analysis indicating that HG downregulated GLUT1 and GLUT4, whereas TXNIP knockdown significantly restored their expression. N Western blot analysis of MAPK signaling showing HG-induced phosphorylation of p38 and JNK, with TXNIP knockdown attenuating these effects. O Treatment with the Trx-1 inhibitor PX-12 abrogated the GPX4 upregulation induced by TXNIP silencing, supporting the requirement of Trx-1 activity in TXNIP-mediated ferroptosis regulation. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Equal amounts of protein were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, IPVH00010), blocked with 5% BSA, and incubated overnight at 4 °C with primary antibodies against TXNIP (ab188865, Abcam, RRID:AB_2923036), Trx-1 (A4024, ABclonal, RRID:AB_2863174), GPX4 (ab125066, Abcam, RRID:AB_10973901), and β-actin (ab115777, Abcam, RRID:AB_10899528).

Techniques: Activation Assay, Protein-Protein interactions, Quantitative RT-PCR, Western Blot, Expressing, Immunoprecipitation, Knockdown, Activity Assay, Phospho-proteomics