Journal: Journal of Translational Autoimmunity

Article Title: Anti-IgLON5 encephalitis is associated with anti-retinal immunological reactivity without retinal alteration

doi: 10.1016/j.jtauto.2026.100359

Figure Lengend Snippet: Indirect immunofluorescence on sections of monkey retina. The fluorescence observed is identified by a black arrow. (A) Patient 4 (IgG4); (B) Patient 1 (IgG4); (C) Patient 3 (IgG1); (D) Patient 5 (IgG4); (E) Patient 2 (IgG1); (F) Patient 1 (IgG4) after immunoadsorption of IgLON5 antibodies; (G) Control with macular edema (IgG1); (H) Control with anti-Hu encephalitis (IgG1); (I) Control with CAR syndrome (IgG4). The different layers of the retina are identified by their initials: pigment epithelium (pe), photoreceptor layer (pr), outer grain layer (og), outer plexiform layer (op), inner grain layer (ig), inner plexiform layer (ip), ganglion cell layer (gc), nerve fibre layer (nf).

Article Snippet: The reactivity of the patients’ sera and/or CSF against the retina was evaluated retrospectively with frozen samples (−80°, EXPLAINEUR biobank) through an indirect immunofluorescence technique using sections of monkey retina (Ref. FA1172-1005, Euroimmun), detected with an FITC-labelled secondary antibody anti-human IgAGM (Euroimmun conjugate) or directed against IgA (ref. F0204, DAKO), IgM (ref. F0203, DAKO), IgG1 (ref. 9052-02, Southern Biotech) and IgG4 (ref. 9200-02, Southern Biotech).

Techniques: Immunofluorescence, Fluorescence, Control

Journal: Journal of Translational Autoimmunity

Article Title: Anti-IgLON5 encephalitis is associated with anti-retinal immunological reactivity without retinal alteration

doi: 10.1016/j.jtauto.2026.100359

Figure Lengend Snippet: Indirect immunofluorescence on sections of monkey retina. The fluorescence observed is identified by a black arrow. (A) Patient 4 (IgG4); (B) Patient 1 (IgG4); (C) Patient 3 (IgG1); (D) Patient 5 (IgG4); (E) Patient 2 (IgG1); (F) Patient 1 (IgG4) after immunoadsorption of IgLON5 antibodies; (G) Control with macular edema (IgG1); (H) Control with anti-Hu encephalitis (IgG1); (I) Control with CAR syndrome (IgG4). The different layers of the retina are identified by their initials: pigment epithelium (pe), photoreceptor layer (pr), outer grain layer (og), outer plexiform layer (op), inner grain layer (ig), inner plexiform layer (ip), ganglion cell layer (gc), nerve fibre layer (nf).

Article Snippet: The reactivity of the patients’ sera and/or CSF against the retina was evaluated retrospectively with frozen samples (−80°, EXPLAINEUR biobank) through an indirect immunofluorescence technique using sections of monkey retina (Ref. FA1172-1005, Euroimmun), detected with an FITC-labelled secondary antibody anti-human IgAGM (Euroimmun conjugate) or directed against IgA (ref. F0204, DAKO), IgM (ref. F0203, DAKO), IgG1 (ref. 9052-02, Southern Biotech) and IgG4 (ref. 9200-02, Southern Biotech).

Techniques: Immunofluorescence, Fluorescence, Control

Journal: Science Advances

Article Title: In vivo membrane engineering traps Gd-based MRI contrast agents for detecting microhepatocellular carcinoma

doi: 10.1126/sciadv.aec9913

Figure Lengend Snippet: ( A ) Representative Western blot analysis of GPC3 expression in HepG2, WRL-68, and Hepa1-6 cells ( n = 3 independent experiments). ( B ) Quantitation of relative GPC3 protein level from (A). Data are presented as means ± SD ( n = 3). Statistical analysis was performed using one-way ANOVA with a Tukey’s post hoc test. ( C ) Flow cytometry analysis of surface GPC3 expression in HepG2, WRL-68, and Hepa1-6 cells. ( D ) Representative IHC images of GPC3 expression (brown) in tumor tissues from orthotopic Hepa1-6 tumor-bearing mice. Scale bar, 50 μm. ( E ) CLSM images of HepG2 and Hepa1-6 cells treated with SPD1 or SPD2 nanoparticles (50 μM; red fluorescence) for 6 hours. Scale bars, 20 μm. ( F ) Time-dependent CLSM imaging of HepG2 cells treated with SPD1 nanoparticles (50 μM) showing membrane-localized fibrillar transformation. Scale bars, 20 μm. ( G ) CLSM analysis of HepG2 cells sequentially incubated with SPD1 or SPD2 nanoparticles (50 μM, 6 hours; red) and FITC-labeled anti-GPC3 antibody (green; 1:200; Abcam, #ab207080). Colocalization (yellow) indicates specific binding of SPD1 to membrane-bound GPC3. Fluorescence intensity and colocalization were quantified using MATLAB. Data are presented as means ± SD ( n = 3); n.s., not significant (one-way ANOVA with Tukey’s post hoc test). Scale bars, 20 μm. ( H ) SEM images of untreated HepG2 and WRL-68 cells or incubated with SPD1 or SPD2 nanoparticles (50 μM, 6 hours). Magnified insets highlight membrane-associated fibrillar structures. ( I ) TEM images of untreated HepG2 cells (top) and those treated with SPD1 nanoparticles (50 μM, 24 hours; bottom). Red arrows indicate membrane-associated nanofibers. Scale bars, 500 nm. ( J ) SEM images showing the persistence of SPD1-derived fibrillar networks on HepG2 cells at 6, 24, and 72 hours posttreatment (50 μM). Scale bars, 2 μm. All experiments were independently repeated three times with consistent and reproducible results.

Article Snippet: After three PBS washes (5 min each), cells were incubated with an FITC-conjugated goat anti-mouse IgG secondary antibody (1:1000; Abbkine, A23210) for 1 hour at room temperature in the dark.

Techniques: Western Blot, Expressing, Quantitation Assay, Flow Cytometry, Fluorescence, Imaging, Membrane, Transformation Assay, Incubation, Labeling, Binding Assay, Derivative Assay

Journal: Science Advances

Article Title: In vivo membrane engineering traps Gd-based MRI contrast agents for detecting microhepatocellular carcinoma

doi: 10.1126/sciadv.aec9913

Figure Lengend Snippet: ( A ) Representative CLSM images of HepG2 cells incubated with SPD1 nanoparticles (red, 50 μM) for 6 hours, followed by treatment with FITC-N 3 (10 to 50 μM, green) for an additional 6 hours. Merged yellow fluorescence indicates successful copper-free click conjugation between DBCO and N 3 on the cell membrane. Cells treated with SPD2+FITC-N 3 (50 μM) served as the nontargeted controls, showing minimal colocalization. Scale bars, 20 μm. ( B ) Time-dependent kinetics of bioorthogonal conjugation quantified by BCA protein assay and ICP-MS. HepG2 cells were pretreated with SPD1 or SPD2 nanoparticles (50 μM, 6 hours), followed by incubation with Gd-DOTA-N 3 (50 μM) for 0.5, 1, 6, or 12 hours. Cells treated with Gd-DOTA-N 3 alone served as baseline controls. ( C ) T 1 -weighted MR images of HepG2 cells treated with Gd-DOTA (50 μM), Gd-DOTA-N 3 (50 μM), or sequentially with SPD1 or SPD2 (50 μM, 6 hours) followed by Gd-DOTA-N 3 (50 μM, 6 hours). ( D ) Quantitative r 1 relaxivity under the corresponding treatment conditions in (C). ( E ) r 1 relaxivity of HepG2 cells preblocked with anti-GPC3 antibody (5 μg/ml, 12 hours; Abcam, #ab207080) before SPD1 treatment (50 μM, 6 hours) followed by Gd-DOTA-N 3 (50 μM, 6 hours). ( F to H ) Cell viability of WRL-68 (F), HepG2 (G), and Hepa1-6 (H) cells after sequential treatment with SPD1 or SPD2 for 6 hours followed by Gd-DOTA-N 3 (50 μM, 6 hours). Cell viability was quantified using the CCK-8 assay. Data are presented as means ± SD { n = 3 for [(A) to (E)]; n = 6 for [(F) to (H)]}. Statistical significance was performed using one-way ANOVA followed by Tukey’s post hoc test. P < 0.05 was considered statistically significant; n.s., not significant. All experiments were independently repeated three times with consistent results.

Article Snippet: After three PBS washes (5 min each), cells were incubated with an FITC-conjugated goat anti-mouse IgG secondary antibody (1:1000; Abbkine, A23210) for 1 hour at room temperature in the dark.

Techniques: Incubation, Fluorescence, Conjugation Assay, Membrane, Bicinchoninic Acid Protein Assay, CCK-8 Assay