Journal: Neural Regeneration Research

Article Title: Human neural stem cell–derived extracellular vesicles protect against ischemic stroke by activating the PI3K/AKT/mTOR pathway

doi: 10.4103/NRR.NRR-D-23-01144

Figure Lengend Snippet: Characterization of hNSCs and hNSC-EVs. (A) Morphology of first- and third-passage hNSCs. Scale bar: 100 μm. (B) The hNSCs were positive for neural stem cell markers. Representative images of hNSCs showing PAX6 in purple, Nestin in purple, and DAPI (nuclear counterstain) in blue. Scale bar: 50 μm. (C) Analysis of NSC multipotentiality via a standard differentiation assay showing upregulation of neuronal and neuroglial markers. Representative images of hNSCs showing βIII-tubulin (neuron) in white, GFAP (neuroglia) in green, and DAPI (nuclear counterstain) in blue. Scale bar: 50 μm. (D) Nanoparticle tracking analysis showing the diameters of the hNSC-EVs. (E) TEM showing hNSC-EV morphology. Scale bar: 100 nm. (F) Immunoblot for CD9, CD63, TSG101, and calnexin expression by hNSC-EVs. (G) hNSC-EV internalization by HUVECs, mouse hippocampal neuronal cells (HT22), and mouse microglial cells (BV2). Representative confocal microscope images of cells with green-labeled cytoplasmic and DAPI-stained nuclei (blue) 6 hours after exposure to PKH26-labeled hNSC-EVs (red). Scale bar: 5 μm. All experiments were performed at least three times. DAPI: 4′,6-Diamidino-2-phenylindole; GFAP: glial fibrillary acidic protein; hNSC: human neural stem cell; hNSC-EVs: hNSC-derived extracellular vesicles; HUVEC: human umbilical vein endothelial cells; NTA: nanoparticle tracking analysis.

Article Snippet: And then, the cells were incubated with the primary antibodies rabbit anti-glial fibrillary acidic protein (GFAP; 1:500, Cat# ab7260, Abcam, Cambridge, MA, USA), rabbit anti-βIII-tubulin (1:100, Cat# ab18207, Abcam), rabbit anti-NeuN (1:80, Abcam, Cat# ab177487) rabbit anti-PAX6 (1:350, Cat# ab195045, Abcam), or rabbit anti-Nestin (1:200, Cat# ab105389, Abcam) overnight at 4°C.

Techniques: Differentiation Assay, Western Blot, Expressing, Microscopy, Labeling, Staining, Derivative Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Interneuron migration impairment and brain region-specific DNA damage response following irradiation during early neurogenesis in mice

doi: 10.1007/s00018-025-05643-7

Figure Lengend Snippet: List of primers used for qRT-PCR

Article Snippet: The following primary antibodies and respective dilutions were used: mouse anti-γH2AX (1:200, Merck Millipore, 05-636), rabbit anti-53BP1 (1:1000, Novus Biologicals, NB100-304), rabbit anti-PH3 (1:500, Cell Signaling, 3377), rabbit anti-CC3 (1:200, Cell Signaling, 9661), mouse anti-Emx1 (1:300, Santa Cruz, sc-398115), rabbit anti-TTF1 (Nkx2.1, 1:250, Abcam, ab76013), mouse anti-Ascl1 (1:500, Santa Cruz, sc-374104), rabbit anti-Pax6 (1:300, BioLegend, 901301) and mouse anti-Nestin (1:300, Invitrogen, MA1-110).

Techniques:

Journal: Brain Structure & Function

Article Title: The Hippo effector TEAD1 regulates postnatal murine cerebellar development

doi: 10.1007/s00429-025-02903-x

Figure Lengend Snippet: Details of all antibodies used in this study for IF analyses

Article Snippet: PAX6 , Rabbit , BioLegend , 901,301 , 1/400.

Techniques:

Journal: Brain Structure & Function

Article Title: The Hippo effector TEAD1 regulates postnatal murine cerebellar development

doi: 10.1007/s00429-025-02903-x

Figure Lengend Snippet: Abnormal migration of developing granule neurons in the absence of Tead1 . Mid-sagittal sections of control ( A-C ) and cKO ( D-F ) cerebella at P20, revealing the expression of PAX6 (green); nuclei were labelled with DAPI (grey). In control cerebella, PAX6 expression was predominantly confined to the IGL, with relatively few PAX6-expressing cells evident within the ML (arrowheads in A - C ). The dotted lines demarcate the boundary between the PCL and the IGL. In comparison, many more PAX6-expressing cells were evident within the ML of cKO cerebella (arrows in D - F , quantified in G ). Furthermore, the width of the ML was significantly reduced in cKO mice ( H ). Scale bar (in F) represents 20 µm for all panels. ns, not significant, * p < 0.05, ** p < 0.01; one-way ANOVA

Article Snippet: PAX6 , Rabbit , BioLegend , 901,301 , 1/400.

Techniques: Migration, Control, Expressing, Comparison

Journal: bioRxiv

Article Title: Increased Reproducibility of Brain Organoids through Controlled Fluid Dynamics

doi: 10.1101/2025.03.05.641438

Figure Lengend Snippet: (A) Brightfield images of day 20 organoids cultured in the RC system, showing uniform morphology and size distribution. Scale bars: 1 mm. (B) Immunofluorescence images of day 20 organoids, stained for PAX6 (green), DCX (white), and DAPI (blue). The main panel provides an overview of the organoid structure, while high-magnification insets, highlighted by a dashed-line rectangle, show the distribution of PAX6+ progenitors and DCX+ migrating neurons. Scale bars: 100 μm. (C) Schematic representation of dorsal and ventral forebrain markers, illustrating marker expression patterns in organoids. (D) Immunofluorescence staining at days 25, 45, and 90, showing expression of PAX6, FOXG1, and SOX2 (green), DCX (white), and TBR1, SATB2, and MAP2 (red) across different time points. Scale bars: 100 μm.

Article Snippet: The following antibodies were used to perform immunostainings: PAX6 antibody (901301; Biolegend; 1/500); FOXG1 antibody (ab18259; abcam; 1/200); SOX2 antibody (ab171380; abcam; 1/200); Doublecortin (E-6) antibody (sc-271390; Santa Cruz; 1/100); TBR1 antibody (ab31940; abcam; 1/200); SATB2 antibody (MA5-32788; Invitrogen; 1:100); MAP2 antibody (M4403; Sigma; 1/500); NKX2.1/TTF-1 antibody (ab133737; abcam; 1/200); DLX2 antibody (sc-393879; Santa Cruz; 1/200).

Techniques: Cell Culture, Immunofluorescence, Staining, Marker, Expressing

Journal: bioRxiv

Article Title: Increased Reproducibility of Brain Organoids through Controlled Fluid Dynamics

doi: 10.1101/2025.03.05.641438

Figure Lengend Snippet: (A) Schematic representation of the experimental workflow. RC organoids from two independent iPSC lines were harvested at day 90, dissociated into single cells, and subjected to single-cell RNA sequencing. (B) UMAP representation of the fetal dorsal cortex (left) ( Polioudakis et al ., 2019 ) and RC organoids (right), illustrating the clustering of cell populations. The pie charts indicate the proportional distribution of different cell types in each dataset. (C) Feature plots displaying the expression patterns of key neural progenitor and neuronal markers, including SOX2 (neural progenitors), PAX6 (dorsal progenitors), DCX (migrating neuroblasts), TUBB3 (neuronal marker), S100B (radial glia and oligodendrocyte progenitors), and NKX2.1 (ventral forebrain marker). (D) UMAP representation of the RC organoid dataset, separated by iPSC lines (RC line 1 and RC line 2), demonstrating consistency between replicates. (E) Proportional representation of cell types across the two RC organoid lines, visualized through dot plots and pie charts, confirming the reproducibility of the differentiation process. Cell types include: ExM_U (maturing excitatory upper enriched), ExDp1 (excitatory deep layer 1), ExM (maturing excitatory), ExN (migrating excitatory), IP (intermediate progenitors), Immature_Ns (immature neurons), InMGE (interneuron medial ganglionic eminence), InCGE (interneuron caudal ganglionic eminence), PgG2M (cycling progenitors in G2/M phase), PgS (cycling progenitors in S phase), vRG (ventricular radial glia), oRG (outer radial glia), RGs (radial glial populations), Prog_mixed (mixed progenitors), OPC (oligodendrocyte progenitor cells), End (Endothelial cells) and Per (pericytes).

Article Snippet: The following antibodies were used to perform immunostainings: PAX6 antibody (901301; Biolegend; 1/500); FOXG1 antibody (ab18259; abcam; 1/200); SOX2 antibody (ab171380; abcam; 1/200); Doublecortin (E-6) antibody (sc-271390; Santa Cruz; 1/100); TBR1 antibody (ab31940; abcam; 1/200); SATB2 antibody (MA5-32788; Invitrogen; 1:100); MAP2 antibody (M4403; Sigma; 1/500); NKX2.1/TTF-1 antibody (ab133737; abcam; 1/200); DLX2 antibody (sc-393879; Santa Cruz; 1/200).

Techniques: RNA Sequencing, Expressing, Marker