Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enhancement of antibiotic therapy in osteomyelitis by inhibiting talin-1-mediated bacterial internalization

doi: 10.3389/fcimb.2026.1823065

Figure Lengend Snippet: Intracellular bacteria impair BMSCs osteogenesis despite antibiotic treatment. (A) Micro-CT analysis showing 3D overview and 2D cross-sectional views of femoral bones from the indicated experimental groups at 30 days post-intervention. (B) Bone-related parameters, including bone mineral density (BMD), bone volume/total volume (BV/TV), trabecular separation (Tb.Sp), trabecular thickness (Tb.Th), cortical porosity (Ct.Po), and trabecular number (Tb.N), were quantified (n = 6). (C, D) Immunofluorescence staining of BMP-2 in the femoral bone tissues of mice, with subsequent quantification (original magnification ×63; scale bar = 50 µm). (E, F) Immunofluorescence staining of Runx-2 in the femoral bones, along with quantitative analysis. (G) Histological analysis of bone tissue sections stained for bacteria in the control, bacterial, and antibiotic-treated groups. Cell nuclei are stained blue with DAPI, cell membranes are stained green with wheat germ agglutinin, and S. aureus is stained red. In the merged image, triangles indicate intracellular bacteria, while pentagrams denote extracellular bacteria. (H) Quantitative assessment of intracellular and extracellular bacteria and their corresponding MOI.

Article Snippet: Antibodies used: BMP-2 (A0231, ABclonal, China), Runx-2 (A11753, ABclonal, China), Alp (A0514, ABclonal, China), Collagen-1 (A21059, ABclonal, China), Talin-1 (14168-1-AP, Proteintech, China), FAK (66258-1-Ig, Proteintech, China), Pan phospho-Serine/Threonine Mouse mAb (AP1067, ABclonal, China), Pan phospho-Tyrosine Mouse mAb (AP0973, ABclonal, China), Mprip (D8G8R, Cell Signaling Technology, USA).

Techniques: Bacteria, Micro-CT, Immunofluorescence, Staining, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enhancement of antibiotic therapy in osteomyelitis by inhibiting talin-1-mediated bacterial internalization

doi: 10.3389/fcimb.2026.1823065

Figure Lengend Snippet: Transcriptome sequencing demonstrated increased global phosphorylation. (A, B) Live/dead cell staining and quantification following bacterial infection at different MOIs (100, 200, 400) and time points (2 h, 4 h) (scale bar = 50 µm; n = 3). (C, D) ALP staining and fluorescence intensity quantification before and after bacterial infection. (E) RT-qPCR analysis of BMP-2, Runx-2, Alp, and Collagen I expression in control and bacteria groups. (F, G) Western blotting and corresponding quantitative analysis of BMP-2, Runx-2, Alp and Collagen I in control and bacteria groups. (H) Volcano plot of transcriptome sequencing differential expression between the control group and bacteria-infected group [Dashed box: phosphorylation-related genes (Top10)]. (I) Heatmap of differential gene expression. (J) Top 20 keggenriched gene pathways based on transcriptome sequencing between the control group and bacteria-infected group (Red font: Extracellular matrix and adhesion-related pathways). (K) GSEA plot of the focal adhesion signaling pathway. (L, M) Western blot analysis and quantitative measurement of Fak protein expression in the control and bacteria-infected groups (n = 3). (N, O) Western blot analysis and quantitative measurement of global phosphotyrosine levels in the control and bacteria-infected groups. (P, Q) Western blot analysis and quantitative measurement of global phosphoserine/threonine levels in the control and bacteria-infected groups.

Article Snippet: Antibodies used: BMP-2 (A0231, ABclonal, China), Runx-2 (A11753, ABclonal, China), Alp (A0514, ABclonal, China), Collagen-1 (A21059, ABclonal, China), Talin-1 (14168-1-AP, Proteintech, China), FAK (66258-1-Ig, Proteintech, China), Pan phospho-Serine/Threonine Mouse mAb (AP1067, ABclonal, China), Pan phospho-Tyrosine Mouse mAb (AP0973, ABclonal, China), Mprip (D8G8R, Cell Signaling Technology, USA).

Techniques: Sequencing, Phospho-proteomics, Staining, Infection, Fluorescence, Quantitative RT-PCR, Expressing, Control, Bacteria, Western Blot, Quantitative Proteomics, Gene Expression

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enhancement of antibiotic therapy in osteomyelitis by inhibiting talin-1-mediated bacterial internalization

doi: 10.3389/fcimb.2026.1823065

Figure Lengend Snippet: Integrated omics reveals talin-1 as key for bacterial internalization. (A) PCA analysis of proteomics and phosphoproteomics. (B) Top 20 significantly enriched go pathways shared (Red font: extracellular matrix and adhesion-related pathways). (C) Volcano plot of proteomics data (No significant difference in Talin-1 protein expression). (D) Volcano plot of phosphoproteomics data (all significantly altered Talin-1 modification sites are up-regulated). (E, F) Western blotting and quantitative analysis of Talin-1 transfection experiments. (G, H) Comparison of intracellular colony formation at 1/3/7 days: si-Talin vs. Bacterial groups (n = 3). (I, J) ALP fluorescence staining and quantitative analysis of fluorescence intensity at 1/3/7 days between si-Talin and bacterial groups. (K) RT-qPCR analysis of BMP-2, Runx-2, Alp, and Collagen I expression in control, bacteria and si-Talin-1 groups. (L, M) Western blotting and corresponding quantitative analysis of BMP-2, Runx-2, Alp and Collagen I in control, bacteria, and si-Talin-1 groups.

Article Snippet: Antibodies used: BMP-2 (A0231, ABclonal, China), Runx-2 (A11753, ABclonal, China), Alp (A0514, ABclonal, China), Collagen-1 (A21059, ABclonal, China), Talin-1 (14168-1-AP, Proteintech, China), FAK (66258-1-Ig, Proteintech, China), Pan phospho-Serine/Threonine Mouse mAb (AP1067, ABclonal, China), Pan phospho-Tyrosine Mouse mAb (AP0973, ABclonal, China), Mprip (D8G8R, Cell Signaling Technology, USA).

Techniques: Phospho-proteomics, Expressing, Modification, Western Blot, Transfection, Comparison, Fluorescence, Staining, Quantitative RT-PCR, Control, Bacteria

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enhancement of antibiotic therapy in osteomyelitis by inhibiting talin-1-mediated bacterial internalization

doi: 10.3389/fcimb.2026.1823065

Figure Lengend Snippet: Mprip physically interacts with Talin-1 to trigger its subsequent phosphorylation. (A) Ranked list of Talin-1 interacting proteins. (B) Confocal fluorescence images showing colocalization of host Mprip and Talin-1 upon S. aureus infection. Blue indicates DAPI; green, Mprip protein; red, Talin-1 protein. (scale bar = 50 µm). (C) Co-IP assay demonstrating the interaction between Talin and Mprip. (D) Transfection efficiency of siRNA targeting Mprip (si-Mprip). (E) Quantitative analysis of si-Mprip transfection (n = 3). (F, G) Western blot analysis showing that knockdown of Mprip decreases the protein expression level of phosphorylated Talin. (H) RT-qPCR analysis of osteogenic markers BMP-2, Runx-2, Alp, and Collagen I in control, bacterial, and si-Mprip groups. (I, J) Western blot analysis and quantification of BMP-2, Runx-2, Alp, and Collagen I expression in control, bacterial, and si-Mprip groups (n = 3).

Article Snippet: Antibodies used: BMP-2 (A0231, ABclonal, China), Runx-2 (A11753, ABclonal, China), Alp (A0514, ABclonal, China), Collagen-1 (A21059, ABclonal, China), Talin-1 (14168-1-AP, Proteintech, China), FAK (66258-1-Ig, Proteintech, China), Pan phospho-Serine/Threonine Mouse mAb (AP1067, ABclonal, China), Pan phospho-Tyrosine Mouse mAb (AP0973, ABclonal, China), Mprip (D8G8R, Cell Signaling Technology, USA).

Techniques: Phospho-proteomics, Fluorescence, Infection, Co-Immunoprecipitation Assay, Transfection, Western Blot, Knockdown, Expressing, Quantitative RT-PCR, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enhancement of antibiotic therapy in osteomyelitis by inhibiting talin-1-mediated bacterial internalization

doi: 10.3389/fcimb.2026.1823065

Figure Lengend Snippet: Talin-1 inhibits osteogenesis by activating the PI3K-AKT pathway. (A, B) Western blot analysis and quantitative comparison of PI3K–AKT activator (740 Y-P at a concentration of 20 μM) and inhibitor (PI3K/AKT-IN-1 (2.62 µM)) treatments. (C, D) Western blot analysis and quantitative measurement of PI3K/p-PI3K and AKT/p-AKT in Control, Bacteria, and Bacteria + si-Talin-1 groups. (E) RT-qPCR analysis of BMP-2, Runx-2, Alp, and Collagen I expression in Control, Bacteria, and Bacteria + PI3K (-) groups, PI3K (-): PI3K/AKT pathway inhibitor. (F, G) Western blot analysis and quantification of BMP-2, Runx-2, Alp, and Collagen I in Control, Bacteria, and Bacteria + PI3K (-) groups. (H) RT-qPCR analysis of BMP-2 and Runx-2 in Control, Bacteria, Bacteria + si-Talin-1, and Bacteria + si-Talin-1 + PI3K(+) groups. (I, J) Western blot analysis and quantitative assessment of BMP-2 and Runx-2 in Control, Bacteria, Bacteria + si-Talin-1, and Bacteria + si-Talin-1 + PI3K (+) groups (n = 3).

Article Snippet: Antibodies used: BMP-2 (A0231, ABclonal, China), Runx-2 (A11753, ABclonal, China), Alp (A0514, ABclonal, China), Collagen-1 (A21059, ABclonal, China), Talin-1 (14168-1-AP, Proteintech, China), FAK (66258-1-Ig, Proteintech, China), Pan phospho-Serine/Threonine Mouse mAb (AP1067, ABclonal, China), Pan phospho-Tyrosine Mouse mAb (AP0973, ABclonal, China), Mprip (D8G8R, Cell Signaling Technology, USA).

Techniques: Western Blot, Comparison, Concentration Assay, Control, Bacteria, Quantitative RT-PCR, Expressing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enhancement of antibiotic therapy in osteomyelitis by inhibiting talin-1-mediated bacterial internalization

doi: 10.3389/fcimb.2026.1823065

Figure Lengend Snippet: UM-164 blocks bacterial internalization and promotes bone formation by inhibiting talin-1 phosphorylation. (A) Virtual screening of small-molecule compounds binding to Talin-1. (B) Molecular docking simulation of UM-164 with Talin-1. (C) SPR assay evaluating the interaction (KD: dissociation constant, which reflects the affinity of the analyte to the target. The smaller the value, the stronger the affinity; Ka: association rate constant, which represents the speed of intermolecular binding. The larger the value, the faster the binding; Kd: dissociation rate constant, which represents the speed of intermolecular dissociation. The larger the value, the faster the dissociation). (D) CCK-8 cell viability assay performed on cells treated with UM-164 at concentrations of 0, 0.001, 0.01, 0.1, 1 and 10 μM. Cell viability decreased significantly as the concentration increased from 0.1 μM to 1 μM. (E) UM-164 treatment decreased the binding affinity between talin-1 and Mprip, as revealed by Co-IP assay. (F, G) Western blot analysis showing that UM-164 treatment decreases the phosphorylation level of Talin-1. (H, I) Intracellular bacterial CFU assays and statistical analysis comparing untreated and UM-164-treated cells (0.1 μM; labeled as “UM-164” in panel I) at 1/3/7 days post-infection. (J, K) RT-qPCR analysis of BMP-2 and Runx-2 expression levels following treatment with different concentrations of UM-164 (0, 0.001, 0.01, 0.1, 1 and 10 μM). (L–N) Western blotting and quantitative analysis of BMP-2 and Runx-2 protein levels after treatment with varying concentrations of UM-164 (0, 0.001, 0.01, 0.1, 1and 10 μM) (n = 3).

Article Snippet: Antibodies used: BMP-2 (A0231, ABclonal, China), Runx-2 (A11753, ABclonal, China), Alp (A0514, ABclonal, China), Collagen-1 (A21059, ABclonal, China), Talin-1 (14168-1-AP, Proteintech, China), FAK (66258-1-Ig, Proteintech, China), Pan phospho-Serine/Threonine Mouse mAb (AP1067, ABclonal, China), Pan phospho-Tyrosine Mouse mAb (AP0973, ABclonal, China), Mprip (D8G8R, Cell Signaling Technology, USA).

Techniques: Phospho-proteomics, Binding Assay, SPR Assay, CCK-8 Assay, Viability Assay, Concentration Assay, Co-Immunoprecipitation Assay, Western Blot, Labeling, Infection, Quantitative RT-PCR, Expressing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Enhancement of antibiotic therapy in osteomyelitis by inhibiting talin-1-mediated bacterial internalization

doi: 10.3389/fcimb.2026.1823065

Figure Lengend Snippet: UM-164 blocks bacterial uptake and restores bone formation in vivo . (A) Micro-CT imaging of femurs from the four experimental groups: control, bacterial infection, antibiotic treatment, and antibiotic + drug treatment. (B) quantitative analysis of micro-CT derived trabecular and cortical bone parameters: BMD, BV/TV, Tb.Sp, Tb.Th, Ct.Po, and Tb.N. (C, D) Immunofluorescence staining of BMP-2 in the femur, along with quantitative analysis (original magnification ×63; scale bar = 50 µm; n = 6). (E, F) Immunofluorescence staining of Runx-2 in the femur, along with quantitative analysis. (G) Histological staining of S. aureus in the femurs of mice across different groups: normal, bacterial infection, antibiotic treatment, and antibiotic + UM-164 treatment. (H) Quantitative analysis of intra- and extracellular bacterial loads, as well as intra- and extracellular bacteria-to-host cell ratios.

Article Snippet: Antibodies used: BMP-2 (A0231, ABclonal, China), Runx-2 (A11753, ABclonal, China), Alp (A0514, ABclonal, China), Collagen-1 (A21059, ABclonal, China), Talin-1 (14168-1-AP, Proteintech, China), FAK (66258-1-Ig, Proteintech, China), Pan phospho-Serine/Threonine Mouse mAb (AP1067, ABclonal, China), Pan phospho-Tyrosine Mouse mAb (AP0973, ABclonal, China), Mprip (D8G8R, Cell Signaling Technology, USA).

Techniques: In Vivo, Micro-CT, Imaging, Control, Infection, Derivative Assay, Immunofluorescence, Staining, Bacteria