Journal: Frontiers in Immunology

Article Title: Unveiling potential diagnostic biomarkers for rheumatoid arthritis through integrated gene expression analysis

doi: 10.3389/fimmu.2026.1645257

Figure Lengend Snippet: Development and validation of the RA diagnostic columnar line graph model. (A) Columnar line graphs used to predict RA incidence. (B) ROC curve analysis to evaluate the clinical utility of the columnar line graph model. (C) DCA curve analysis to assess the clinical utility of the columnar line graph model. (D) Box plot illustrating the expression levels of GABARAPL1, FKBP5, MREG, PCDH9, and SLAMF8.

Article Snippet: Subsequently, they were incubated with the primary antibodies against PCDH9 (Abcam, ab233710), GABARAPL1 (Abcam, ab109364), FKBP5 (Proteintech, 14155-1-AP), SLAMF8 (Novus Biologicals, AF1907), and GAPDH (Proteintech, 10494-1-AP) for an overnight period at 4°C.

Techniques: Biomarker Discovery, Diagnostic Assay, Expressing

Journal: Frontiers in Immunology

Article Title: Unveiling potential diagnostic biomarkers for rheumatoid arthritis through integrated gene expression analysis

doi: 10.3389/fimmu.2026.1645257

Figure Lengend Snippet: Analysis of hub gene expression in synovial tissues from RA patients. (A) Representative MRI scans illustrating synovial thickening. (B) H&E staining of synovial tissue in trauma control (TC) and RA samples (n = 3). (C) Comparative analysis of mRNA expression levels for GABARAPL1, FKBP5, MREG, PCDH9, and SLAMF8 in TC and RA samples (n = 9). (D) Quantitative assessment of protein concentrations for GABARAPL1, FKBP5, PCDH9, and SLAMF8 in TC and RA samples (n = 3, the samples derive from the same experiment and that gels/blots were processed in parallel. original blots/gels are presented in Supplementary Figure 1 ). (E) IF staining results showed protein expression of GABARAPL1, FKBP5, PCDH9, and SLAMF8 in TC and RA samples, visualized with DAPI (blue) and specific antibodies (green) (n = 3). The means ± SD are used to represent the data, while ns (not significant), * P < 0.05, ** P < 0.01, and * P < 0.001 indicate significance levels.

Article Snippet: Subsequently, they were incubated with the primary antibodies against PCDH9 (Abcam, ab233710), GABARAPL1 (Abcam, ab109364), FKBP5 (Proteintech, 14155-1-AP), SLAMF8 (Novus Biologicals, AF1907), and GAPDH (Proteintech, 10494-1-AP) for an overnight period at 4°C.

Techniques: Gene Expression, Staining, Control, Expressing

Journal: Frontiers in Immunology

Article Title: Unveiling potential diagnostic biomarkers for rheumatoid arthritis through integrated gene expression analysis

doi: 10.3389/fimmu.2026.1645257

Figure Lengend Snippet: Correlation between candidate gene expression and inflammatory indices in the RA cohort. Scatter plots show the associations between PCR-measured expression levels of the candidate genes (GABARAPL1, FKBP5, PCDH9, and SLAMF8) and serum inflammatory markers (ESR and CRP) within RA patients only (n = 9). Spearman’s rank correlation was used to estimate the correlation coefficient ( R ), with p values adjusted for multiple testing using the Benjamini–Hochberg false discovery rate (FDR).

Article Snippet: Subsequently, they were incubated with the primary antibodies against PCDH9 (Abcam, ab233710), GABARAPL1 (Abcam, ab109364), FKBP5 (Proteintech, 14155-1-AP), SLAMF8 (Novus Biologicals, AF1907), and GAPDH (Proteintech, 10494-1-AP) for an overnight period at 4°C.

Techniques: Gene Expression, Expressing

Journal: Scientific Reports

Article Title: FKBP51 disrupts the insulin signaling pathway and impairs mitochondrial bioenergetics in HepG2 cells

doi: 10.1038/s41598-026-40414-9

Figure Lengend Snippet: FKBP51 overexpression dampens insulin signaling in HepG2 cells but does not hinder its effects on glucose metabolism. HepG2 cells were transfected with either a plasmid coding Myc-DDK-tagged human FKBP51 (FKBP51) or an empty pCMV6 plasmid (Control) and were stimulated for 0.5 h with 100 nM insulin. ( A ) Representative blot ( n = 8), ( B ) FKBP51 protein levels, ( C ) Akt phosphorylation, ( D ) P70S6K phosphorylation, ( E ) FOXO1, ( F and G ) Glucose production assay ( n = 4), ( H ) mRNA levels (G6P, PCK1 y PDK4) ( n = 4), ( I ) Representative blot ( n = 3), GSK3β phosphorylation, ( J ) Representative imagens of glycogen synthesis assay and ( K ) Glycogen synthesis ( n = 4). One-way analysis of variance (ANOVA) followed by Tukey post hoc test. * p < 0.05 compared with control and square brackets with FKBP51 with insulin. Data are shown as means ± SEM.

Article Snippet: Proteins were resolved in SDS-polyacrylamide gels and subjected to immunoblotting overnight using antibodies specific for FKBP51 (1:1000, 12210 S, Cell Signaling, Massachusetts, USA), Akt (1:2,000, 2920 S, Cell Signaling), phospho-Ser 473 AKT (1:1000, 9271 S, Cell Signaling), phospho-p70S6K Thr389 (1:1000, 9206 S, Cell Signaling), p70S6K (1:2,000, 90205 S, Cell Signaling), phospho-FOXO1 Ser256 (1:1000, 9461 S, Cell Signaling), FOXO1 (1:1000, 14952 S, Cell Signaling), phospho-DRP1 Ser637 (1:1000, ab193216, Abcam, Cambridge, UK), DRP1 (1:1000, ab184247, Abcam), OPA1 (1:1000, ab157457, Abcam), Mfn1 (1:1000, 14739 S, Cell Signaling), Mfn2 (1:1000, 9482 S, Cell Signaling), mt-HSP70 (1:1000, MA3-D28, Invitrogen, Waltham, MA, USA), GAPDH (1:1000, 97166 S, Cell Signaling), OXPHOS (1:1000 ab110413, Abcam), phospo-GSK-3β Ser9 (1:1000, 5558, Cell Signaling) and GSK-3β (1:1,1000, 9832, Cell Signaling).

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Phospho-proteomics

Journal: Scientific Reports

Article Title: FKBP51 disrupts the insulin signaling pathway and impairs mitochondrial bioenergetics in HepG2 cells

doi: 10.1038/s41598-026-40414-9

Figure Lengend Snippet: FKBP51 is partially localized in mitochondria in HepG2 cells, but does not alter mitochondrial morphology. HepG2 cells were transfected with either a plasmid coding Myc-DDK-tagged human FKBP51 (FKBP51) or an empty pCMV6 plasmid (Control) and were stimulated for 3 h with 100 nM insulin and 200 nM CCCP for 2 h. ( A ) Representative image of immunofluorescence confocal microscopy, mtHSP70 (red) for mitochondria, FKBP51 (green). Segment lines represent the cellular contours. Scale bar: 24 μm, ( B ) Quantification of mitochondria (mtHSP70)-FKBP51 colocalization using Mander’s coefficient for cell imagens, ( C ) Quantification of FKBP51-mitochondria (mtHSP70) colocalization using Mander’s coefficient for cell imagens ( n = 6), ( D ) Representative blot mitochondria-cytosol subcellular fractionation ( n = 4), ( E ) Representative image of immunofluorescence confocal microscopy MitoTracker Green FM (200 nM for 0.2 h) in live-cells, ( F ) Quantification of average mitochondrial area, ( G ) Quantification of the of mitochondria number per cell in images cells and ( H ) Mean individual mitochondrial volume of HepG2 cells ( n = 5), ( I ) Representative image of Transmission Electron Microscopy, ( J ) Quantification of mitochondrial area, ( K ) Quantification of mitochondrial perimeter, ( L ) Quantification of mitochondrial aspect ratio. One-way analysis of variance (ANOVA) followed by Tukey post hoc test. * p < 0.05 compared with control. Data are shown as means ± SEM.

Article Snippet: Proteins were resolved in SDS-polyacrylamide gels and subjected to immunoblotting overnight using antibodies specific for FKBP51 (1:1000, 12210 S, Cell Signaling, Massachusetts, USA), Akt (1:2,000, 2920 S, Cell Signaling), phospho-Ser 473 AKT (1:1000, 9271 S, Cell Signaling), phospho-p70S6K Thr389 (1:1000, 9206 S, Cell Signaling), p70S6K (1:2,000, 90205 S, Cell Signaling), phospho-FOXO1 Ser256 (1:1000, 9461 S, Cell Signaling), FOXO1 (1:1000, 14952 S, Cell Signaling), phospho-DRP1 Ser637 (1:1000, ab193216, Abcam, Cambridge, UK), DRP1 (1:1000, ab184247, Abcam), OPA1 (1:1000, ab157457, Abcam), Mfn1 (1:1000, 14739 S, Cell Signaling), Mfn2 (1:1000, 9482 S, Cell Signaling), mt-HSP70 (1:1000, MA3-D28, Invitrogen, Waltham, MA, USA), GAPDH (1:1000, 97166 S, Cell Signaling), OXPHOS (1:1000 ab110413, Abcam), phospo-GSK-3β Ser9 (1:1000, 5558, Cell Signaling) and GSK-3β (1:1,1000, 9832, Cell Signaling).

Techniques: Transfection, Plasmid Preparation, Control, Immunofluorescence, Confocal Microscopy, Fractionation, Transmission Assay, Electron Microscopy

Journal: Scientific Reports

Article Title: FKBP51 disrupts the insulin signaling pathway and impairs mitochondrial bioenergetics in HepG2 cells

doi: 10.1038/s41598-026-40414-9

Figure Lengend Snippet: FKBP51 overexpression decreases Mitofusin 2 protein levels in HepG2 cells. Cells were transfected with either a plasmid coding Myc-DDK-tagged human FKBP51 (FKBP51) or an empty pCMV6 plasmid (Control) and were stimulated for 3 h with 100 nM insulin, mtHSP70 was used as loading control. ( A ) Representative blot of proteins of mitochondrial dynamics ( n = 10), ( B ) Mitofusin 1 protein levels (MFN1), ( C ) Mitofusin 2 protein levels (MFN2), ( E ) Ratio LOPA/SOPA1, ( D ) Dynamin-related protein 1 (Drp1) phosphorylation, ( F ) Mitochondrial fission protein 1 (Fis1 protein levels), ( G ) Dynamin-related protein 1 (Drp1), protein levels, ( H ) Representative blot of mitochondrial oxidative phosphorylation chain (OXPHOS), ( I ) NADH: ubiquinone oxidoreductase subunit B8 (NDUFB8, complex I) protein level, J Succinate dehydrogenase iron-sulfur subunit (SDHB, complex II), ( K ) Ubiquinol-cytochome c reductase core protein 2 (UQCRC2, complex III), ( L ) Cytochrome c oxidase subuni1 (MTCO1, complex IV) and ( M ) ATP synthase subunit alpha (ATP5A, complex V) ( n = 8). One-way analysis of variance (ANOVA) followed by Tukey post hoc test. * p < 0.05 compared with control. Data are shown as means ± SEM.

Article Snippet: Proteins were resolved in SDS-polyacrylamide gels and subjected to immunoblotting overnight using antibodies specific for FKBP51 (1:1000, 12210 S, Cell Signaling, Massachusetts, USA), Akt (1:2,000, 2920 S, Cell Signaling), phospho-Ser 473 AKT (1:1000, 9271 S, Cell Signaling), phospho-p70S6K Thr389 (1:1000, 9206 S, Cell Signaling), p70S6K (1:2,000, 90205 S, Cell Signaling), phospho-FOXO1 Ser256 (1:1000, 9461 S, Cell Signaling), FOXO1 (1:1000, 14952 S, Cell Signaling), phospho-DRP1 Ser637 (1:1000, ab193216, Abcam, Cambridge, UK), DRP1 (1:1000, ab184247, Abcam), OPA1 (1:1000, ab157457, Abcam), Mfn1 (1:1000, 14739 S, Cell Signaling), Mfn2 (1:1000, 9482 S, Cell Signaling), mt-HSP70 (1:1000, MA3-D28, Invitrogen, Waltham, MA, USA), GAPDH (1:1000, 97166 S, Cell Signaling), OXPHOS (1:1000 ab110413, Abcam), phospo-GSK-3β Ser9 (1:1000, 5558, Cell Signaling) and GSK-3β (1:1,1000, 9832, Cell Signaling).

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Phospho-proteomics

Journal: Scientific Reports

Article Title: FKBP51 disrupts the insulin signaling pathway and impairs mitochondrial bioenergetics in HepG2 cells

doi: 10.1038/s41598-026-40414-9

Figure Lengend Snippet: FKBP51 overexpression impairs mitochondrial bioenergetics. HepG2 cells were transfected with either a plasmid coding Myc-DDK-tagged human FKBP51 (FKBP51) or an empty pCMV6 plasmid (Control) and were stimulated for 3 h with 100 nM insulin. ( A ) Oxygen consumption rate (OCR) of cell, measured sequentially for 5 min, (CCCP 100 nM) ( n = 5), ( B ) Representative image of confocal microscopy of HepG2 cells loaded with TMRM then treated with CCCP 100 nM to dissipate the mitochondrial transmembrane potential. ΔB-C (ΔBasal-CCCP) was calculated as the mean fluorescence at 60 s before CCCP addition minus mean fluorescence during the last of the last 60 s of the recording, ( C ) Quantification of Δbasal-CCCP fluorescence ( n = 8), ( D ) Representative image of confocal microscopy MitoSOX 5 µM for 0.2 h with CCCP 200 nM for 1 h positive control, ( E ) Quantification of MitoSOX fluorescence ( n = 6), ( F ) Intracellular ATP levels of cells, ( G ) Graphical representation of the movement of calcium into the mitochondria by the Rhod-2 AM (4 µM for 0.2 h) probe in response to histamine 100 mM, ( H ) Area under the curve (AUC) of Ca 2+ movement to the mitochondria by the Rhod-2 AM probe in response to histamine and Slope the first 10 s in response to histamine by the Rhod-2 probe ( n = 4), ( I ) Graphical representation of the cytosolic Ca 2+ by the Fluo-4 AM (4.4 µM for 0.2 h) probe in response to histamine 100 mM, ( J ) Area under the curve (AUC) of Ca 2+ movement to the mitochondria by the Fluo-4 AM probe in response to histamine and Slope the first 10 s in response to histamine by the Fluo-4 AM probe ( n = 5). ( K ) proposed model. One-way analysis of variance (ANOVA) followed by Tukey post hoc test. * p < 0.05 compared with control and square brackets compared with FKBP51 with insulin. Data are shown as means + SEM.

Article Snippet: Proteins were resolved in SDS-polyacrylamide gels and subjected to immunoblotting overnight using antibodies specific for FKBP51 (1:1000, 12210 S, Cell Signaling, Massachusetts, USA), Akt (1:2,000, 2920 S, Cell Signaling), phospho-Ser 473 AKT (1:1000, 9271 S, Cell Signaling), phospho-p70S6K Thr389 (1:1000, 9206 S, Cell Signaling), p70S6K (1:2,000, 90205 S, Cell Signaling), phospho-FOXO1 Ser256 (1:1000, 9461 S, Cell Signaling), FOXO1 (1:1000, 14952 S, Cell Signaling), phospho-DRP1 Ser637 (1:1000, ab193216, Abcam, Cambridge, UK), DRP1 (1:1000, ab184247, Abcam), OPA1 (1:1000, ab157457, Abcam), Mfn1 (1:1000, 14739 S, Cell Signaling), Mfn2 (1:1000, 9482 S, Cell Signaling), mt-HSP70 (1:1000, MA3-D28, Invitrogen, Waltham, MA, USA), GAPDH (1:1000, 97166 S, Cell Signaling), OXPHOS (1:1000 ab110413, Abcam), phospo-GSK-3β Ser9 (1:1000, 5558, Cell Signaling) and GSK-3β (1:1,1000, 9832, Cell Signaling).

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Confocal Microscopy, Fluorescence, Positive Control