Journal: Fluids and barriers of the CNS

Article Title: Neurogenesis and glial impairments in congenital hydrocephalus: insights from a BioGlue-induced fetal lamb model.

doi: 10.1186/s12987-025-00630-3

Figure Lengend Snippet: Fig. 3 Expressions and distributions of Sox2, Vimentin, Aq4, MOBP, MOG and MBP in HCP. A Sox2 gene and protein expressions were higher in HCP compared to control group at E105. However, as gestation progressed, its expression notably decreased. B Upregulation of Vimentin and Aq4 relative expressions were seen, particularly after E125. Western blot analysis showed significantly higher Vimentin protein level in the hydrocephalic groups at E140. C MOBP and MOG relative expressions upregulated at E105 but after E125, their gene expressions and MBP protein level decreased significantly (Data in the bar graphs represent mean ± SEM, green bars: control, blue bars: HCP) (*p < 0.05, **p < 0.01). D Immunostainings of Sox2 (green), Vimentin (red), MBP (magenta) and DAPI in control and hydrocephalic brains at E105, E125 and E140 (× 20, scale bar 50 μm)

Article Snippet: Blots were treated with primary antibodies against Pax6 (Abcam, ab5790, 1:1000), Sox2 (NovusBio, NB110-37235, 1:2000), GFAP (NovusBio, NBP1-05198, 1:5000), Vimentin (Sigma, V6630, 1:200), Olig2 (Millipore, ABE1024, 1:1000), MBP (AvesLab, MBP89927985, 1:2000) and DCX (Invitrogen, PA5-17428, 1:1000) overnight at + 4 °C, followed by a one-hour treatment with secondary antibodies (anti-rabbit IgG HRP-linked, Cell Signaling Technology, 7074S, 1:1000; anti-mouse IgG HRP-linked, Cell Signaling Technology, 7076S, 1:1000; donkey anti-chicken IgY H&L (HRP), Abcam, ab16349, 1:2000 and goat anti-Guinea pig IgG H&L (HRP), Abcam, ab97155, 1:10000) at room temperature.

Techniques: Control, Expressing, Western Blot

Journal: Fluids and barriers of the CNS

Article Title: Neurogenesis and glial impairments in congenital hydrocephalus: insights from a BioGlue-induced fetal lamb model.

doi: 10.1186/s12987-025-00630-3

Figure Lengend Snippet: Fig. 6 Alterations in neurogenesis and gliogenesis in fetal-onset hydrocephalus. During normal early neurodevelopment, neuroepithelial cells (NECs) in the neural tube differentiate into multipotent apical radial glial cells (aRGs) in the VZ. Subsequently, asymmetric divisions of aRGs generate basal progenitors (BPs) in the SVZ, including basal radial glial cells (bRGs) and basal intermediate progenitors (bIPs), which contribute to the production of cortical neurons and macroglia, essential for the development of the neocortex and formation of cortical folds. In this research on hydrocephalus in fetal lambs, disruptions in the VZ were observed as early as E105. Initial compensatory mechanisms occurred through increased proliferation of the NSCs and NPCs, followed by decreases in important neurogenic regulators such as Pax6 and Sox2, particularly at term. Notably, this surge was associated with significant upregulation of DCX, suggesting active neurogenesis at this early stage. However, this proliferation was accompanied by a decline in the expression of critical neurogenic markers such as Pax6 and Sox2, particularly noticeable towards the term, indicating compromised neuronal repair capabilities. Additionally, alterations in the expression of Ascl1 and Tbr2 pointed to disrupted neuronal differentiation pathways, with fluctuating expression patterns evident across various gestational stages. The progression of hydrocephalus was also marked by increased expression of astroglial markers (GFAP, Vimentin, and Aq4), along with pronounced myelin damage indicated by the reduced expressions of MOBP, MOG, and Sox10 as the gestation advanced. Collectively, these findings suggest that surgical intervention between E105 and E125 may offer a critical therapeutic window, providing a potentially effective period for prenatal surgery to mitigate the irreversible damage due to hydrocephalus (created on BioRender.com)

Article Snippet: Blots were treated with primary antibodies against Pax6 (Abcam, ab5790, 1:1000), Sox2 (NovusBio, NB110-37235, 1:2000), GFAP (NovusBio, NBP1-05198, 1:5000), Vimentin (Sigma, V6630, 1:200), Olig2 (Millipore, ABE1024, 1:1000), MBP (AvesLab, MBP89927985, 1:2000) and DCX (Invitrogen, PA5-17428, 1:1000) overnight at + 4 °C, followed by a one-hour treatment with secondary antibodies (anti-rabbit IgG HRP-linked, Cell Signaling Technology, 7074S, 1:1000; anti-mouse IgG HRP-linked, Cell Signaling Technology, 7076S, 1:1000; donkey anti-chicken IgY H&L (HRP), Abcam, ab16349, 1:2000 and goat anti-Guinea pig IgG H&L (HRP), Abcam, ab97155, 1:10000) at room temperature.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Macrophages activate mesenchymal stem cells to acquire cancer-associated fibroblast-like features resulting in gastric epithelial cell lesions and malignant transformation in vitro

doi: 10.3892/ol.2018.9703

Figure Lengend Snippet: Gastric epithelial cells incubated with CM from macrophage-hucMSCs exhibit several properties of stem cells. (A) GES-1 cells were incubated with CM from hucMSCs or macrophage-hucMSCs for 48 h, and then seeded in non-adherent culture conditions for spheroid formation. Depicted are representative images of spheroid colonies after 15 days. Magnification, ×400; scale bar, 50 µm. (B) Quantification of the soft agar colonies shown in (A). (C) Western blot analysis of Nanog, SOX2, and BMI-1 in GES-1 cells (the controls) and GES-1 cells co-cultured with CM from hucMSCs and macrophage-hucMSCs after 48 h. The expression of all analyzed proteins is significantly enhanced in the GES-1 cells cultured with CM from macrophage-hucMSCs, compared with the control. (D) Representative image of single-colony formations of GES-1 cells incubated with CM from hucMSCs or CM from macrophage-hucMSCs. (E) The number of colonies is depicted as mean ± standard deviation. *P<0.05, # P<0.01; GES-1 cells treated with medium only served as the control (magnification, ×100). Macrophage-hucMSC, human umbilical cord-derived mesenchymal stem cells pre-cultured with macrophages for 48 h; CM, conditioned medium; SOX2, SRY-box 2; BMI-1, polycomb complex protein BMI-1.

Article Snippet: The membranes were incubated with 5% skimmed milk to block non-specific protein at room temperature for 1 h. Membranes were incubated overnight at 4°C with primary antibodies at a dilution of 1:800 for rabbit polyclonal anti-N-cadherin (cat. no. BS2224; Bioworld Technology, Inc., St. Louis Park, MN, USA), 1:1,000 for rabbit polyclonal anti-E-cadherin (cat. no. BS1098; Bioworld Technology, Inc.), 1:500 for rabbit polyclonal anti-vimentin (cat. no. BS1855; Bioworld Technology, Inc.), 1:500 for rabbit polyclonal anti-α-SMA (cat. no. BS8796; Bioworld Technology, Inc.), 1:800 for rabbit polyclonal anti-B-cell lymphoma-2 (Bcl-2) (cat. no. BS70205; Bioworld Technology, Inc.), 1:500 for rabbit polyclonal anti-Bcl-2-associated X (Bax) (cat. no. BS1030; Bioworld Technology, Inc.), 1:500 for rabbit polyclonal anti-SOX2 (cat. no. BS6161; Bioworld Technology, Inc.), 1:1,000 for rabbit polyclonal anti-Bmi-1 (cat. no. BS6015; Bioworld Technology, Inc.), 1:10,000 for rabbit polyclonal anti-β-actin (cat. no. AP0060; Bioworld Technology, Inc.), 1:1,000 for mouse monoclonal anti-FAP (cat. no. sc-71094; Santa Cruz Biotechnology, Inc.,) and 1:1,000 for rabbit polyclonal anti-Nanog (cat. no. sc-33759; Santa Cruz Biotechnology, Inc.).

Techniques: Incubation, Western Blot, Cell Culture, Expressing, Standard Deviation, Derivative Assay