Journal: Frontiers in Immunology

Article Title: A cell line–derived, immune-competent neurospheroid model to study neuroinflammation and human brain disorders

doi: 10.3389/fimmu.2026.1792896

Figure Lengend Snippet: SH-SY5Y and U138 MG cell differentiation into neuron-like (NLCs) and astrocyte-like cells (ALCs). (A, B) Representative immunofluorescence images of β3-tubulin (in yellow) and Nestin (in green) for undifferentiated SH-SY5Y cells (Ctrl) and NLCs differentiated on plastic (Diff.) or on Geltrex (Diff. on Geltrex). Nuclei were counterstained with DAPI (blue). (C, D) Quantification of the fluorescence intensity for β3-tubulin and Nestin normalized to DAPI signals. (E) Relative mRNA expression levels of the proneural transcription factors Mash1 , Nurr1 , and NeuroD1 in undifferentiated and differentiated SH-SY5Y cells. (F) Representative S100B (in red) immunofluorescence images for undifferentiated U138 MG cells or ALCs differentiated on plastic (Diff.) or on Geltrex (Diff. on Geltrex). (G, H) Quantification of the U138 MG proliferating fractions in differentiated and control conditions measuring the EdU-incorporating cells. (I) Quantification of the S100B fluorescence intensity normalized to DAPI signals. Data are shown as mean ± SD from a minimum of three independent replicates (n ≥ 3). Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. *p< 0.03; **p< 0.02; ***p< 0.0002; ****p< 0.0001. Scale bar: 100μm. FI, Fluorescence intensity; Mash1, Mammalian achaete-scute homolog 1; NeuroD1, Neuronal differentiation 1; Nurr1, Nuclear receptor related 1 protein.

Article Snippet: β3-Tubulin , 2G10 , Mouse , sc-80005 , Santa Cruz Biotechnology, Inc., Dallas, TX, USA , 2 μg/mL.

Techniques: Cell Differentiation, Immunofluorescence, Fluorescence, Expressing, Control

Journal: Frontiers in Immunology

Article Title: A cell line–derived, immune-competent neurospheroid model to study neuroinflammation and human brain disorders

doi: 10.3389/fimmu.2026.1792896

Figure Lengend Snippet: Formation and morphological assessment of bi-hNSPHs. (A) Representative bright-field microscopy images showing the formation and growth of bi-hNSPHs within 14 days. Scale bar: 200 µm. (B) Growth curves based on the measurement of bi-hNSPHs areas (µm²) and diameters (µm) every other day. (C) H&E staining of bi-hNSPHs cryosection at 14 days to reveal their internal cellular disposition. Scale bar: 100 µm. (C) Representative immunofluorescence images of neuronal markers for SH-SY5Y – β3-tubulin in yellow, Synapsin in red, MAP2 in magenta and Nestin in green – and astrocytic markers for U138 MG – S100B in red. (D) MAP2, Microtubule associated protein 2.

Article Snippet: β3-Tubulin , 2G10 , Mouse , sc-80005 , Santa Cruz Biotechnology, Inc., Dallas, TX, USA , 2 μg/mL.

Techniques: Microscopy, Staining, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: A cell line–derived, immune-competent neurospheroid model to study neuroinflammation and human brain disorders

doi: 10.3389/fimmu.2026.1792896

Figure Lengend Snippet: Internal composition of tri-hNSPHs. (A) Representative immunofluorescence images of tri-hNSPH cryosections at 4, 7, 9, 11, and 14 days of culture. Sections were stained for the astrocyte marker S100B (in red), the neuronal marker β3-tubulin (in yellow), and the microglia/macrophage marker Iba-1 (in cyan). Nuclei were counterstained with DAPI (blue). (B) Representative immuno-fluorescence images of neuronal – Synapsin in red, MAP2 in magenta and Nestin in green – and microglial – HLA-DR in red – markers on cryosectioned tri-hNSPHs. Scale bar: 50-100 µm. Iba-1, Ionized calcium-binding adapter molecule 1; MAP2, Microtubule associated protein 2.

Article Snippet: β3-Tubulin , 2G10 , Mouse , sc-80005 , Santa Cruz Biotechnology, Inc., Dallas, TX, USA , 2 μg/mL.

Techniques: Immunofluorescence, Staining, Marker, Fluorescence, Binding Assay

Journal: Cell Reports Medicine

Article Title: Blood-brain barrier-penetrative lipid nanoparticles enable systemic delivery of TRIM11 mRNA to disaggregate Tau in Alzheimer’s disease models

doi: 10.1016/j.xcrm.2026.102685

Figure Lengend Snippet: PLNP-mRNA exhibits enhanced BBB penetration and robust gene transfection in vitro and in vivo (A) Schematic representation of features of PLNP-mRNA. (B) Flow cytometry analysis of cellular uptake of Cy5-labeled PLNP-mRNA in SH-SY5Y cells after 4-h incubation. (C) Corresponding confocal fluorescence images of intracellular Cy5 signal. Scale bars, 10 μm ( n = 3). (D) GFP mRNA transfection efficiency of PLNPs in SH-SY5Y cells after 24-h incubation as determined by flow cytometry and confocal microscopy (inset). Scale bars, 50 μm ( n = 3). (E) In vivo whole-brain fluorescence imaging (left) and (right) quantification of Cy5-labeled PLNP-mRNA distribution in C57BL/6 mice 0.5 h post-intravenous injection (1 mg Cy5-mRNA equiv./kg), showing enhanced brain accumulation relative to controls ( n = 3). (F and G) (F) Ex vivo fluorescence imaging of brains isolated from treated mice and (G) quantification of Cy5 fluorescence intensity specifically in the hippocampus. Scale bars: 1 mm (left) and 300 μm (right) ( n = 3). (H) Representative confocal images showing co-localization of Cy5 signal with neurons (β3-Tubulin), astrocytes (GFAP), and microglia (Iba-1). Scale bars, 50 μm ( n = 3). (I and J) (I) Quantification of Cy5 fluorescence associated with each cell type and (J) fold change in Cy5-positive cell populations ( n = 3 independent experiments). (K) Schematic showing PMPC-mediated targeting of PLNP-mRNA to the BBB via binding to chTs and nAChRs on endothelial cells. (L) Brain-wide GFP expression in C57BL/6 mice 24 h after systemic administration of PLNP-GFP mRNA, visualized by ex vivo fluorescence imaging. Scale bars: 1 mm (left) and 300 μm (right). (M and N) (M) Quantitative analysis of GFP fluorescence and (N) fold-change in GFP-positive cells in the hippocampal region, compared with LNP-mRNA controls ( n = 3). (O) Representative images showing GFP expression and phosphorylated Tau (AT8) in 3×Tg-AD mice following PLNP-mGFP administration. Scale bars, 150 μm. (P) Quantification of GFP fluorescence intensity in treated mice ( n = 3). Data are presented as mean ± SD. For in vitro studies ( B–2D), mRNA concentration was 2 μg/mL. For in vivo experiments ( E–2P), dosage was 1 mg mRNA equiv./kg.

Article Snippet: β3-Tubulin Monoclonal Antibody , HUABIO , Cat#ET1602-4;RRID: AB_2938592.

Techniques: Transfection, In Vitro, In Vivo, Flow Cytometry, Labeling, Incubation, Fluorescence, Confocal Microscopy, Imaging, Injection, Ex Vivo, Isolation, Binding Assay, Expressing, Concentration Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Modulation of Wnt/β‐Catenin Pathway by Aesculus hippocastanum Extract Enhances Temozolomide Sensitivity in Glioblastoma Cells

doi: 10.1111/jcmm.70979

Figure Lengend Snippet: HCE and TMZ combination inhibits Wnt/β‐catenin signalling and modulates glioma stemness and differentiation markers in U87 cells. U87 cells were treated with HCE (500 μg/mL), TMZ (100 μM), or their combination, for 48 h. Western blot analysis of β‐catenin and Wnt‐1 was performed (A, B; A1, B1). ELISA kit for phospho‐GSK‐3β (Ser9) and total GSK‐3β was shown in (C). Nestin (D–H) and β3‐tubulin (I–M) were assessed by IF. Images of IF were captured at 40× magnification. Data are representative of three independent experiments. One‐Way ANOVA. (A1) * p < 0.05 versus Control; *** p < 0.001 versus Control; # p < 0.05 versus TMZ. (B1) * p < 0.05 versus Control; *** p < 0.001 versus Control; ## p < 0.01 versus TMZ. (C) * p < 0.05 versus Control; *** p < 0.001 versus Control; # p < 0.05 versus TMZ. (H) ** p < 0.01 versus Control; *** p < 0.001 versus Control; ## p < 0.01 versus TMZ. (M) ** p < 0.01 versus Control; *** p < 0.001 versus Control; ## p < 0.01 versus TMZ.

Article Snippet: The following primary antibodies were used: Nestin (1:100, Arigo Laboratories, Cat. No. ARG52345 ), β‐3 tubulin (1:100, Santa Cruz Biotechnology, Dallas, TX, USA; sc‐80005), Bax (1:100, Invitrogen; Cat. No. MA5‐14003), and p53 (1:100, Santa Cruz Biotechnology, Dallas, TX, USA; sc‐126).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Modulation of Wnt/β‐Catenin Pathway by Aesculus hippocastanum Extract Enhances Temozolomide Sensitivity in Glioblastoma Cells

doi: 10.1111/jcmm.70979

Figure Lengend Snippet: Effects of HCE and TMZ on oxidative and nitrosative stress in U87 cells. U87 cells were treated with HCE (500 μg/mL), TMZ (100 μM), or their combination for 48 h. Levels of ROS (A), ROMO‐1 (B), GSH (C), MDA (D), 3‐NT (E), and NO 2 − (F) were assessed. Data are representative of three independent experiments. One‐Way ANOVA. (A) ** p < 0.01 versus Control; *** p < 0.001 versus Control; # p < 0.05 versus TMZ. (B) * p < 0.05 versus Control; ** p < 0.01 versus Control; *** p < 0.001 versus Control; # p < 0.05 versus TMZ. (C) *** p < 0.001 versus Control; # p < 0.05 versus TMZ. (D) *** p < 0.001 versus Control; ### p < 0.001 versus TMZ. (E) *** p < 0.001 versus Control; # p < 0.05 versus TMZ. (F) ** p < 0.01 versus Control; *** p < 0.001 versus Control; ### p < 0.001 versus TMZ.

Article Snippet: The following primary antibodies were used: Nestin (1:100, Arigo Laboratories, Cat. No. ARG52345 ), β‐3 tubulin (1:100, Santa Cruz Biotechnology, Dallas, TX, USA; sc‐80005), Bax (1:100, Invitrogen; Cat. No. MA5‐14003), and p53 (1:100, Santa Cruz Biotechnology, Dallas, TX, USA; sc‐126).

Techniques: Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Modulation of Wnt/β‐Catenin Pathway by Aesculus hippocastanum Extract Enhances Temozolomide Sensitivity in Glioblastoma Cells

doi: 10.1111/jcmm.70979

Figure Lengend Snippet: Effects of HCE and TMZ on U87 spheroid area over time. U87 spheroids were treated with HCE (500 μg/mL), TMZ (100 μM), or their combination, and spheroid area was assessed at Day 1 (A–D), Day 3 (E–H), and Day 7 (I–L, score M). Data are representative of three independent experiments. Two‐Way ANOVA. (M) * p < 0.05 versus Control.

Article Snippet: The following primary antibodies were used: Nestin (1:100, Arigo Laboratories, Cat. No. ARG52345 ), β‐3 tubulin (1:100, Santa Cruz Biotechnology, Dallas, TX, USA; sc‐80005), Bax (1:100, Invitrogen; Cat. No. MA5‐14003), and p53 (1:100, Santa Cruz Biotechnology, Dallas, TX, USA; sc‐126).

Techniques: Control