Journal: Translational Psychiatry

Article Title: Astrocytic APOE3-Christchurch expression ameliorates brain amyloid-β pathology in 5xFAD mice

doi: 10.1038/s41398-026-04002-9

Figure Lengend Snippet: ( A - C ) Aβ40 and Aβ42 levels in the soluble TBS fractions ( A ), detergent soluble (TBS-X) fractions ( B ), and insoluble neutralized formic acid (FA) fractions ( C ) of cortical brain homogenates in the 5xFAD mice were measured by ELISA at 8 months of age, normalized to protein concentrations of each fraction. ( D ) Soluble oligomeric Aβ (oAβ) levels in the TBS-X fractions of cortical brain homogenates in the 5xFAD mice were measured by ELISA at 8 months of age, normalized to protein concentrations. ( E, F ) Amounts of full-length APP in the TBS-X fractions of mouse cortical lysates were measured by western blot at 8 months of age, normalized to those of β-actin (ACTB). Data are expressed as means ± SEM (GFP: N = 12; APOE3: N = 18; APOE3Ch: N = 13; Open circles: females, closed circles: males). Injection group differences were analyzed using two-way ANOVA, with Tukey’s multiple comparisons test, adjusting for sex. *, P < 0.05; **, P < 0.01.

Article Snippet: Recombinant Aβ42 monomer (1 mg/ml) (Stressmarq, #SPR-485) was incubated at room temperature for 10 min. To dissolve the peptide, 7.5 μl of cold DMSO (Sigma Aldrich) was added, followed by 92.5 μl of cold PBS (PH 7.4) to reach a final concentration of 220 μM monomeric Aβ42.

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

Journal: Translational Psychiatry

Article Title: Astrocytic APOE3-Christchurch expression ameliorates brain amyloid-β pathology in 5xFAD mice

doi: 10.1038/s41398-026-04002-9

Figure Lengend Snippet: ( A ) Isogenic iPSC-derived astrocytes (iPSC-ACs) with homozygous APOE3 (left) and APOE3Ch (right) were immunostained for GFAP (green) and S100b (red). DAPI (blue) stains nuclei. Scale bars: 100 µm. ( B ) APOE mRNA levels in the iPSC-ACs were measured by RT-qPCR, normalized to those of β-actin ( ACTB ). ( C, D ) Amounts of APOE in the conditioned medium (CM) from iPSC-ACs were measured by western blot, normalized to protein amounts of cell lysates. ( E ) Effects of the CM from iPSC-ACs on Aβ42 aggregation was assessed by western blot using 6E10 antibody. ( F - G ) Populations of Aβ fibrils (F; > 150 kDa), oligomers (G; 37–150 kDa), and monomers ( H ; < 10 kDa) were quantified. Data expressed as means ± SEM (n = 3–5 independent differentiation batches). Group differences were analyzed using two-tailed student t-test or one-way ANOVA with Tukey’s multiple comparisons test. *, P < 0.05.

Article Snippet: Recombinant Aβ42 monomer (1 mg/ml) (Stressmarq, #SPR-485) was incubated at room temperature for 10 min. To dissolve the peptide, 7.5 μl of cold DMSO (Sigma Aldrich) was added, followed by 92.5 μl of cold PBS (PH 7.4) to reach a final concentration of 220 μM monomeric Aβ42.

Techniques: Derivative Assay, Quantitative RT-PCR, Western Blot, Two Tailed Test

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: Patient-derived PanNET tumoroids express HH signaling proteins and respond to HH pathway activation and inhibition. A Combined fluorescence and phase-contrast images of 21-day-old human PanNET tumoroids (PanNET3) stained for chromogranin A (CHGA) and HH proteins (PTCH1, SHH). B Four patient-derived PanNET tumoroid lines (PanNET1–4) were exposed to recombinant human SHH N-terminal peptide (100 ng/mL) or the SMO inhibitor vismodegib (VISMO, 20 µM) for 72 h and changes in mRNA expression were analyzed by RT-qPCR. mRNA changes were normalized to HPRT1 expression and DMSO vehicle control. ( n = 4 unique patient lines). * = p < 0.05, ** = p < 0.01, **** = p < 0.0001 by Two-way ANOVA with Sidak post-test. C EdU labeling showing proliferation of dissociated PanNET3 cells following 5-day exposure to: the HH agonists SHH-N (100 ng/mL) and SAG (10 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 1 µM). D Quantitation of the percentage of EdU-positive PanNET tumor cells following 5-day treatment. ( n = 3 replicates from one patient tumoroid line). * = p < 0.05, by One-way ANOVA with Tukey post-test. E SHH or SAG were co-administered with the respective pharmacologic inhibitors and EdU uptake was evaluated after 7 days. F Immunofluorescent images of CHGA, SHH, and PTCH1 expression in a second PanNET tumoroid line (PanNET5). G , H EdU labeling was evaluated in PanNET5 tumoroids following 7-day treatment with SHH-N (200 ng/mL) and SAG (20 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 5 µM). I Crystal violet staining of human BON-1 PanNET cells after 48 h treatment. J BrdU incorporation in BON-1 cells after 48 h treatment with HH agonists SHH-N and SAG, ( K ) SMO inhibitors vismodegib and sonidegib, and ( L ) inhibitors of GLI1/2 signaling. M Immunofluorescent images of CHGA, SHH, and PTCH1 expression in tumoroids derived from a metastatic ileal NET (IL-NET-met1). N , O EdU labeling was assayed in the IL-NET-met1 tumoroid line following 7-day treatment with the same drug concentrations used for PanNET5. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Derivative Assay, Activation Assay, Inhibition, Fluorescence, Staining, Recombinant, Expressing, Quantitative RT-PCR, Control, Labeling, Quantitation Assay, BrdU Incorporation Assay

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: Loss of Men1 in enteric glial cells stimulates GLI1/2-dependent transcriptional reprogramming. A Combined fluorescence and phase contrast images of 5-day-old primary enteric glial cell (EGC) cultures from Sox10-CreER T2 ; LSL-tdTomato mice and CreER T2 negative controls. Top panel shows TdTomato+ EGCs after 48 h exposure to 4-hydroxytamoxifen 4-OHT (2 µM). B TdTomato + EGCs were sorted by FACS to enrich for a pure SOX10 + cell population. C Combined fluorescence and phase contrast images of FACS-enriched SOX10-tdTomato + EGCs. D Fluctuations in HH pathway mRNA levels were evaluated in SOX10-tdTomato + EGCs 72 h following siRNA-mediated Men1 silencing. siRNA treatment consisted of four pooled siRNAs targeting the Men1 gene ( si -Men1 , 25 nM) or non-targeting (si-NT, 25 nM) controls. ( n = 5). E Immunofluorescence images of SHH expression in si-NT and si-Men1 treated EGCs (SHH = red pseudo-color, DAPI = blue). Inset shows higher power image. F Western blot analysis of si-NT and si-Men1 EGCs after 72 h treatment. SHH-FL = 55 kDa full length peptide; SHH- N = 22 kDa N-terminal peptide. ( n = 3). G Quantitation of protein expression in panel (F) normalized to GAPDH loading control. ( n = 3). H Relative fold-change in glial lineage transcripts and ( I ) neuroendocrine and neural progenitor transcripts in si-NT and si-Men1 treated EGCs. ( n = 6). J qPCR analysis of HH pathway genes and ( K ) neuroendocrine and neural progenitor transcriptsin si-NT and si-Men1 EGCs after 72 h treatment with GANT61 (10 µM) or vismodegib (VISMO 20 µM). ( n = 3). For all plots, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by Two-way ANOVA with Sidak post-test. L Immunofluorescence images of si-NT and si-Men1 EGCs after 96 h siRNA knockdown and 72 h treatment with vehicle or GANT61. Menin = green, GFAP = magenta, SHH = yellow. M Significant GSEA pathways in enteric glial cells following 5-days of si- Men1 knockdown compared to non-targeting control. GSEA was performed on Men1 -depleted cells and cells co-treated with ( N ) si- Men1 , si- Gli1 , and si- Gli2 , and ( O ) cells co-treated with si- Men1 and GANT61 (10 µM). P – R KEGG pathway enrichment analysis comparing the same groups as shown in panels ( M – O ). S Heatmap showing significant DEGs mapped to the cell cycle, HH signaling, epigenetic regulation, neural stem cell (NSC) reprogramming, neuronal and neuroendocrine differentiation

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Fluorescence, Immunofluorescence, Expressing, Western Blot, Quantitation Assay, Control, Knockdown