Journal: Cell Communication and Signaling : CCS

Article Title: Rho GTPases signaling mediates aggressiveness and differentiation in neuroblastoma tumors

doi: 10.1186/s12964-025-02649-3

Figure Lengend Snippet: ARHGAP31 /CdGAP is associated with undifferentiated cells, and its downregulation provokes Cdc42 overexpression. A mRNA expression of ARHGAP31 in adrenergic tumors, mesenchymal tumors or neural crest tissues. Long rank test p value is shown, * = p < 0.05. Dataset: GSE908030. B mRNA expression of ARHGAP31 in SK-N-SH CD44hi-derived undifferentiated secondary tumorspheres (TS2) compared to the bulk cell line. * = p < 0.05, n = 4. C Immunofluorescence showing CdGAP expression in NB48T CD44hi-derived undifferentiated tumorspheres compared to CD44negative-derived ones. D Tumorsphere formation efficiency of SK-N-SH or NB48T PDX-derived cell lines after CdGAP knock down by siRNA or control. * = p < 0.05; *** = p < 0.005. E Expression of the undifferentiated marker nestin in SK-N-SH and NB48T PDX-derived tumorspheres after CdGAP knock down by siRNA or control. ** = p < 0.01; **** = p < 0.001. F Expression of Cdc42 in SK-N-SH and NB48T PDX-derived TS cells after CdGAP knock down by siRNA or control. Representative images from NB48T cells are shown. **** = p < 0.001. G Expression of Sox9 protein in cells from tumorspheres generated from NB48T or sorted CD44hi SK-N-SH and NB48T cells, after CdGAP knock down by siRNA or control. * = p < 0.05; **** = p < 0.001. H Cell viability in CHLA 20 cells, with or without downregulation of CdGAP, over 72 h (** = p < 0.01). I Quantification of proliferating Ki67 expressing cells in NB48T cells with or without downregulation of CdGAP. Graph shows mean ± SEM. Scale bar = 20μm. (* = p < 0.05, n = 3). J Morphological characterization of CHLA 20 and NB48T cells with or without downregulation of CdGAP. Cell segmentation on the cell contour is shown in red. Scale bar = 20 μm. K Immunofluorescence for acetylated tubulin, and DAPI in CHLA 20 cells with or without downregulation of CdGAP. The presence of acetylated tubulin in cell extensions is indicated with red asterisks. Scale bar = 20 μm. L Quantification of the cells positive for acetylated tubulin staining in cellular protrusions after the indicated treatments in CHLA20 cells (mean ± SEM, * = p < 0.05; ** = p < 0.001, n = 3)

Article Snippet: For immunofluorescence, cells were fixed with 4% PFA in PBS for 15 min, permeabilized with 0.2% Triton X-100 in PBS for 10 min at 4 °C and blocked with PBS + 1% BSA for at least 1 h. Primary antibodies used were: CD44 mouse (1:200 BD Pharmingen), Cdc42 rabbit (1:1000 Cell Signaling), Ki67 rabbit (1:200 Thermo Scientific), Nestin rabbit (1:1000 Millipore), Nestin mouse (1:1000 R&D Systems), α-tubulin mouse (1:1000 Sigma), acetylated tubulin mouse (1:1000, Santa Cruz), Tuj1-βIII tubulin mouse (1:1000 abcam) and CdGAP (1:500, Santa Cruz).

Techniques: Over Expression, Expressing, Derivative Assay, Immunofluorescence, Knockdown, Control, Marker, Generated, Staining

Journal: Journal of Stem Cell Research & Therapy

Article Title: No Detection of Potential Cancer Risk for Free-Viral Reprogrammed Stem Cell-Derived Dopaminergic Neurons from Adult Mice Fibroblasts

doi: 10.4172/2157-7633.1000286

Figure Lengend Snippet: Figure 2: Differentiation of neural progenitor cells from iPSCs. Producing neural progenitor cells from iPSCs were manufactured using the Neural Induction Medium during Days 20-35 (Figure 2a and 2b; 2a with nestin antibody staining), and there was no significant alteration in the morphology of the the parallel cultured control group (Figure 2c).

Article Snippet: Mouse Nestin Antibody (R & D system, MAB2736) was applied as a neural progenitor mark for the verification of neuronal progenitor cells.

Techniques: Staining, Cell Culture, Control

Journal: iScience

Article Title: Enhanced differentiation of neural progenitor cells in Alzheimer’s disease into vulnerable immature neurons

doi: 10.1016/j.isci.2025.112446

Figure Lengend Snippet: Enhanced neuronal activity and early differentiation in APPswe cerebral organoids (A–C) Quantification of the perimeter (B) and area (C) of control and APPswe cerebral organoids cultured for one month. n = 80–104, from 3 independent differentiations. Scale bar, 500 μm. (D–F) Neuronal activity was measured by a multi-electrode array in control and APPswe cerebral organoids cultured for one month. n = 11–12, from 3 independent differentiations. (G–I) Immunohistochemistry was performed to analyze the distribution of (H) nesitn- and (I) DCX-positive cells in control and APPswe cerebral organoids cultured for one month. n = 9, from 3 independent differentiations. Scale bar, 20 μm. (J–M) Immunoblotting to determine expression levels of SOX2 (K), DCX (L), and TUJ1 (M) in control and APPswe cerebral organoids cultured for one month. n = 10–14, from 3 independent differentiations. (N–P) Flow cytometry analysis was performed on control and APPswe cerebral organoids cultured for one month to assess the expression levels of (O) nestin- and (P) DCX-positive populations. n = 3 from 3 independent differentiations. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001 (Student’s t test). Error bar ± SEM.

Article Snippet: After blocking, the cells were incubated for 2 hours at 4°C with primary antibodies: a nestin PE-conjugated antibody (R&D Systems) and an antibody against DCX.

Techniques: Activity Assay, Control, Cell Culture, Immunohistochemistry, Western Blot, Expressing, Flow Cytometry

Journal: iScience

Article Title: Enhanced differentiation of neural progenitor cells in Alzheimer’s disease into vulnerable immature neurons

doi: 10.1016/j.isci.2025.112446

Figure Lengend Snippet: Increased mitochondrial ROS and its impact on early differentiation in APPswe NPCs (A–C) Immunocytochemistry analysis of nestin (B) and DCX (C) in control and APPswe NPCs and neurons, differentiated for one week. Scale bar, 40 μm. NPCs, n = 30, from 3 independent differentiations; neuron, n = 75, from 6 independent differentiations. (D) Increased MitoSOX signal in APPswe NPCs. n = 3, from 3 independent differentiations. (E) Reduced utilization of OXPHOS in glucose metabolism in APPswe NPCs. n = 5, from 3 independent differentiations. (F–H) Normalization of increased nestin (G) and decreased DCX (H) expression in APPswe NPCs with EUK8 treatment. Scale bar, 40 μm. n = 75, from 5 independent differentiations. (I) Schematic of the experiment categorizing control NPCs into high-ROS and low-ROS groups based on MitoSOX signal intensity. (J–L) No significant differences in nestin and DCX expression between high-ROS and low-ROS NPC groups. Scale bar, 20 μm. n = 40, from 3 independent differentiations. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 (ANOVA test followed by Dunnett’s post hoc analysis or Student’s t test). Error bar ± SEM.

Article Snippet: After blocking, the cells were incubated for 2 hours at 4°C with primary antibodies: a nestin PE-conjugated antibody (R&D Systems) and an antibody against DCX.

Techniques: Immunocytochemistry, Control, Expressing