Journal: iScience

Article Title: GAL and F2R as immune diagnostic biomarkers for fetal growth restriction

doi: 10.1016/j.isci.2026.115228

Figure Lengend Snippet: Targeted additional analysis of the three key IR-DEGs (A) Chromosomal positions of the key IR-DEGs are presented. (B) A PCA plot illustrates the distribution of samples based on the expression profiles of the 3 key IR-DEGs. The x axis and y axis correspond to the first two principal components (PC1 and PC2), respectively, and the percentage of total variance explained by each component is indicated in parentheses adjacent to the axis labels. (C) Comparative expression levels of three crucial IR-DEGs in FGR, which integrates datasets GSE24129 , GSE100415 , and GSE147776 . (D) ROC curves were used to validate the efficacy of three crucial IR-DEGs in predicting FGR, quantifying the diagnostic performance of each gene for FGR identification. (E) A nomogram for predicting the risk of FGR is constructed based on three IR-DEGs, specifically F2R, GAL, and CXCL10. For each of these genes, a corresponding point value is assigned according to its expression level; the total point score is calculated by summing the individual gene points, and this total score is further converted to the predicted risk of developing FGR. (F) A calibration curve for the nomogram is shown, comparing the nomogram-predicted risk of FGR ( x axis) with the actually observed risk ( y axis). The diagonal line represents an ideal prediction scenario where predicted and observed risks are identical. The dashed line (“Apparent”) denotes the model’s performance before bias correction, while the solid line (“Bias-corrected”) represents performance after bias correction.

Article Snippet: IHC staining was performed according to previously established protocols using primary antibodies against GAL (Bioss, Beijing, China, catalog number: bs-0017M, RRID: AB_10855141) and F2R (Bioss, Beijing, China, catalog number: bs-0828R, RRID: AB_10857704).

Techniques: Expressing, Diagnostic Assay, Construct

Journal: iScience

Article Title: GAL and F2R as immune diagnostic biomarkers for fetal growth restriction

doi: 10.1016/j.isci.2026.115228

Figure Lengend Snippet: An evaluation focusing on immune infiltration related to two IR-DEGs (A) Stacked bar plot showing the relative abundance of 22 immune cell subtype proportions between FGR and AGA samples. (B) A boxplot is employed to visualize the differentiation in ratios of 22 immune cell types, with a specific focus on comparisons between FGR and AGA. (C) A Spearman correlation network is constructed to illustrate the correlative relationships between two IR-DEGs (namely GAL and F2R) and infiltrating immune cells in FGR. This network visualization explicitly displays how each of the four target IR-DEGs correlates with the 22 types of infiltrating immune cells, facilitating intuitive recognition of positive or negative correlation patterns between the genes and immune cell subsets. (D) Expression levels of F2R are significantly higher in FGR tissues ( n = 11) compared to AGA samples ( n = 25). Data are presented as mean ± SEM. (E) Expression levels of GAL are significantly elevated in FGR tissues ( n = 11) relative to AGA specimens ( n = 25). Data are presented as mean ± SEM. (F) qRT-PCR analysis demonstrates that F2R mRNA expression is significantly upregulated in FGR placental tissues compared to AGA controls ( p = 0.0019). Data are presented as mean ± SEM. (G) qRT-PCR analysis reveals a significant downregulation of GAL mRNA in FGR placental tissues compared to AGA controls ( p = 0.0015). Data are presented as mean ± SEM. Additionally, representative images of IHC staining for F2R and GAL in FGR and AGA patients are presented, illustrating both high and low expression levels of the two genes. All staining images are shown at magnifications of ×40 and ×200, with scale bars clearly indicated for reference. Statistical p values were calculated via the chi-square test, where ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: IHC staining was performed according to previously established protocols using primary antibodies against GAL (Bioss, Beijing, China, catalog number: bs-0017M, RRID: AB_10855141) and F2R (Bioss, Beijing, China, catalog number: bs-0828R, RRID: AB_10857704).

Techniques: Construct, Expressing, Quantitative RT-PCR, Immunohistochemistry, Staining

Journal: bioRxiv

Article Title: Winding-Up of Fibrin Fibers as a Novel Mechanism of Platelet-Mediated Fiber Compaction

doi: 10.64898/2026.02.15.705975

Figure Lengend Snippet: Platelets become labelled by fluorescent fibrin fibers during clot retraction A) Time course of unconstrained clot retraction by platelets (4x10 in 400 μl of 12% plasma in PBS) for a total of 60 minutes at room temperature. Experiments have been performed 6x using blood from different donors and typical examples of blood from two different donors are shown (experiments III and IV are repetitions of I and II the day after; although a longer lag-phase is observed, the final retraction volume is similar for the four experiments). B) Unconstrained clot retraction (1x10 platelets per ml in 50% plasma/50% PBS) in presence of fibrinogen-Alexa 488. Image acquisition was started immediately after thrombin addition at a focal plane 100 μm above the bottom of the well and image stacks were collected (61 focal planes, step size 0.5 μm) for a time period of 20:27 min (100 frames with a time interval of 12.5 sec). See also associated video 1. The experiment has been performed twice using platelets of the same donor. Shown are the first, intermediate and last time points of a depth color-coded time-lapse video (left panel; scale bar 10 μm) and the maximal intensity projections (MIPs) of the same time points (right panel; scale bar 10 μm). C) Constrained clot retraction (1x10 platelets per ml in 50% plasma/50% PBS, fibrinogen-Alexa 488 final concentration 12.5 μg/ml) between two holders. Clots were induced by addition of thrombin (2.5U/ml final), fixed at the indicated retraction times, embedded in gelatin, flash frozen and cryosections (14 μm) were stained for the integrin subunit αIIb (magenta; scale bar 5 μm). The time course was performed twice using PRP from two different donors (retraction assays were repeated, although not for all time points, more than eight times using blood from different donors, with consistent results). Image acquisition was performed using a wide-field epi fluorescence microscope (BX41; Olympus) equipped with a Plan 100x/1.25 NA oil objective, a camera (DP70; Olympus), and the acquisition software analySIS (Olympus).

Article Snippet: Reagents: Neutral formalin (Sigma-Aldrich, Saint-Quentin Fallavier, France), Pluronic F-127 (Sigma-Aldrich, P2443), thrombin (Sigma-Aldrich, T4648), Jasp-K4-HA (a HAK-actin probe, kind gift of Paul Guichard/Virginie Hamel), apyrase (Sigma-Aldrich, A6535), heparin (Sigma-Aldrich, H3393), PGI2 (Sigma-Aldrich, P6188) Antibodies: mouse anti-integrin αIIb (Santa-Cruz, sc-7310), monoclonal rabbit anti-HA (Cell Signaling, C29F4), rabbit anti-non muscle myosin II heavy chain (Covance, PRB-440P), rabbit anti-phospho S19 myosin light chain (Abcam, ab2480), monoclonal rabbit anti-myosin light chain 2 (Abcam, ab92721), Cy3 goat anti-mouse IgG (Jackson, 115-165-166), Cy3 goat anti-rabbit IgG (Jackson, 111-165-003), Alexa 647 goat anti-rabbit IgG (Jackson, 111-605-003)

Techniques: Clinical Proteomics, Concentration Assay, Staining, Fluorescence, Microscopy, Software

Journal: bioRxiv

Article Title: Winding-Up of Fibrin Fibers as a Novel Mechanism of Platelet-Mediated Fiber Compaction

doi: 10.64898/2026.02.15.705975

Figure Lengend Snippet: Fibrin fibers are organized in a "cage-like" fashion around platelets within a constrained clot A-D: Two typical examples (out of 39 acquisitions of four experiments using blood from different donors) of platelets in a constrained clot with attached fibrin fibers are shown (1x10 platelets per ml PBS/50% plasma and fibrinogen-Alexa 488). Clot retraction was allowed to take place in an inoculation loop (see methods) for 15 min before fixation and immunofluorescence staining of the integrin subunit αIIb (magenta). Samples were then processed for expansion (scale bars 10 μm = 2.5μm after correction for expansion; indicated z-levels are not corrected for expansion; see also associated animation, video 2). A) Four focal planes from a stack of images (z-level as indicated) showing a platelet in a clot (fibrin fibers in green, plasma membrane staining using an antibody against the integrin subunit αIIb in magenta and the merge). B) 3D reconstruction of the image stack used in A (46 planes, step size 0.5 μm), a bulb is indicated by an arrow. C) Four focal planes show another typical platelet in a clot (z-level as indicated). D) 3D image reconstruction of the image stack used in C (28 planes, step size 1 μm), a bulb is indicated by an arrow.

Article Snippet: Reagents: Neutral formalin (Sigma-Aldrich, Saint-Quentin Fallavier, France), Pluronic F-127 (Sigma-Aldrich, P2443), thrombin (Sigma-Aldrich, T4648), Jasp-K4-HA (a HAK-actin probe, kind gift of Paul Guichard/Virginie Hamel), apyrase (Sigma-Aldrich, A6535), heparin (Sigma-Aldrich, H3393), PGI2 (Sigma-Aldrich, P6188) Antibodies: mouse anti-integrin αIIb (Santa-Cruz, sc-7310), monoclonal rabbit anti-HA (Cell Signaling, C29F4), rabbit anti-non muscle myosin II heavy chain (Covance, PRB-440P), rabbit anti-phospho S19 myosin light chain (Abcam, ab2480), monoclonal rabbit anti-myosin light chain 2 (Abcam, ab92721), Cy3 goat anti-mouse IgG (Jackson, 115-165-166), Cy3 goat anti-rabbit IgG (Jackson, 111-165-003), Alexa 647 goat anti-rabbit IgG (Jackson, 111-605-003)

Techniques: Clinical Proteomics, Immunofluorescence, Staining, Membrane

Journal: bioRxiv

Article Title: Winding-Up of Fibrin Fibers as a Novel Mechanism of Platelet-Mediated Fiber Compaction

doi: 10.64898/2026.02.15.705975

Figure Lengend Snippet: Coiled-up fibrin fibers above spread platelets Two representative examples (out of 18 acquisitions from three independent experiments using blood from different donors) of fibrin fiber accumulations above spread platelets in the 2D fiber-retraction assay (4 μl plasma per ml PBS, see methods section) are shown (see also animation, video 3). Samples were stained for the αIIb integrin subunit and expanded (scale bars 10 μm = 2.5 μm after correction for expansion, indicated z-levels are not corrected for expansion). A) Three different focal planes (z-levels as indicated) from an image stack showing two spread platelets and attached fibrin fibers. B) 3D reconstruction of the image stack used in A (51 planes, step size 0.25 μm), shown are three different view angles. C) Three different focal planes from an image stack illustrating a spread platelet and attached, coiled-up fibrin fibers. D) 3D reconstruction of the image stack used in C (50 planes, step size 0.25 μm), shown are three different view angles. The fibrin fibers are additionally displayed as depth-color coded (Fire LUT) reconstructions to highlight the straight fiber parts along the z-axis. E) Same fiber image stacks as in C and D. Left panels are MIPs of the green fibrin fibers along the z, x and y axes. Right panels are the same projections color coded in the HSB space (Hue, Saturation, Brightness) according to the orientation of individual fiber segments. The fibers are well aligned along the z-axis, while they crisscross horizontally.

Article Snippet: Reagents: Neutral formalin (Sigma-Aldrich, Saint-Quentin Fallavier, France), Pluronic F-127 (Sigma-Aldrich, P2443), thrombin (Sigma-Aldrich, T4648), Jasp-K4-HA (a HAK-actin probe, kind gift of Paul Guichard/Virginie Hamel), apyrase (Sigma-Aldrich, A6535), heparin (Sigma-Aldrich, H3393), PGI2 (Sigma-Aldrich, P6188) Antibodies: mouse anti-integrin αIIb (Santa-Cruz, sc-7310), monoclonal rabbit anti-HA (Cell Signaling, C29F4), rabbit anti-non muscle myosin II heavy chain (Covance, PRB-440P), rabbit anti-phospho S19 myosin light chain (Abcam, ab2480), monoclonal rabbit anti-myosin light chain 2 (Abcam, ab92721), Cy3 goat anti-mouse IgG (Jackson, 115-165-166), Cy3 goat anti-rabbit IgG (Jackson, 111-165-003), Alexa 647 goat anti-rabbit IgG (Jackson, 111-605-003)

Techniques: Clinical Proteomics, Staining

Journal: bioRxiv

Article Title: Winding-Up of Fibrin Fibers as a Novel Mechanism of Platelet-Mediated Fiber Compaction

doi: 10.64898/2026.02.15.705975

Figure Lengend Snippet: Strongly compacted fibrin fibers around the center of platelets in the 2D fiber-retraction assay Two representative examples of platelets are shown with bulbs encircled by fibrin fibers in the 2D fiber-retraction assay (4 μl plasma/ml, see methods section) behaving similar to platelets in a clot (see also animation, video 4). Samples were stained for the αIIb integrin subunit (A, B) or the myosin light chain (MLC; C,D) and processed for expansion (scale bars 10 μm = 2.5 μm after correction for expansion, indicated z-levels are not corrected for expansion). Experiment was repeated 4x with blood from different donors with a total of 18 image stacks acquired for fibrin/integrin and 62 for fibrin/myosin staining. A) Three different focal planes from an image stack showing two platelets, one spread and one with bulbs and attached, compacted fibrin fibers. B) 3D image reconstruction of the image stack used in A (34 planes, step size 0.25 μm), shown are three different view angles. C) Three different focal planes from an image stack illustrating another platelet with bulbs and attached, rolled-up, compacted fibrin fibers. D) 3D image reconstruction of the image stack used in C (55 planes, step size 0.25 μm), shown are three different view angles.

Article Snippet: Reagents: Neutral formalin (Sigma-Aldrich, Saint-Quentin Fallavier, France), Pluronic F-127 (Sigma-Aldrich, P2443), thrombin (Sigma-Aldrich, T4648), Jasp-K4-HA (a HAK-actin probe, kind gift of Paul Guichard/Virginie Hamel), apyrase (Sigma-Aldrich, A6535), heparin (Sigma-Aldrich, H3393), PGI2 (Sigma-Aldrich, P6188) Antibodies: mouse anti-integrin αIIb (Santa-Cruz, sc-7310), monoclonal rabbit anti-HA (Cell Signaling, C29F4), rabbit anti-non muscle myosin II heavy chain (Covance, PRB-440P), rabbit anti-phospho S19 myosin light chain (Abcam, ab2480), monoclonal rabbit anti-myosin light chain 2 (Abcam, ab92721), Cy3 goat anti-mouse IgG (Jackson, 115-165-166), Cy3 goat anti-rabbit IgG (Jackson, 111-165-003), Alexa 647 goat anti-rabbit IgG (Jackson, 111-605-003)

Techniques: Clinical Proteomics, Staining

Journal: bioRxiv

Article Title: Winding-Up of Fibrin Fibers as a Novel Mechanism of Platelet-Mediated Fiber Compaction

doi: 10.64898/2026.02.15.705975

Figure Lengend Snippet: A fibrin rosette is located close to a ring-like actin organization in spread platelets Four representative examples (out of 20 acquisitions, experiment repeated 4x using blood from different donors) of a fibrin rosette associated with spread platelets in the 2D fiber-retraction assay (7 μl plasma per ml PBS, see methods section) are shown (see also animation, video 6). Samples were stained for the αIIb integrin subunit as well as for actin and processed for expansion (scale bars 10 μm = 2.5 μm after correction for expansion). A) 3D image reconstruction of an image stack (30 planes, step size 0.33 μm) of fibrin fibers (green) and integrin or actin staining (upper and lower panels respectively, magenta). B) Similar example as in A (17 planes, step size 0.33 μm). C) Another example of a fibrin rosette with intercalated actin nodules (34 planes, step size 0.33 μm). D) An example showing a long fibrin fiber associated with the fibrin rosette (35 planes, step size 0.38 μm).

Article Snippet: Reagents: Neutral formalin (Sigma-Aldrich, Saint-Quentin Fallavier, France), Pluronic F-127 (Sigma-Aldrich, P2443), thrombin (Sigma-Aldrich, T4648), Jasp-K4-HA (a HAK-actin probe, kind gift of Paul Guichard/Virginie Hamel), apyrase (Sigma-Aldrich, A6535), heparin (Sigma-Aldrich, H3393), PGI2 (Sigma-Aldrich, P6188) Antibodies: mouse anti-integrin αIIb (Santa-Cruz, sc-7310), monoclonal rabbit anti-HA (Cell Signaling, C29F4), rabbit anti-non muscle myosin II heavy chain (Covance, PRB-440P), rabbit anti-phospho S19 myosin light chain (Abcam, ab2480), monoclonal rabbit anti-myosin light chain 2 (Abcam, ab92721), Cy3 goat anti-mouse IgG (Jackson, 115-165-166), Cy3 goat anti-rabbit IgG (Jackson, 111-165-003), Alexa 647 goat anti-rabbit IgG (Jackson, 111-605-003)

Techniques: Clinical Proteomics, Staining