Journal: bioRxiv
Article Title: Spatially precise neuron formation via hydrogel mediated modulation of the host astrocyte response
doi: 10.64898/2025.12.20.695501
Figure Lengend Snippet: A representative image of astrocytes transduced with A, F) AAV-mCherry and B, E, G) AAV-SOX2 and stained with nestin/OCT3-4/Ki67 marker (cyan), mCherry (red), GFAP (yellow), and DAPI (blue). C) Quantification data showing the population of nestin+, mCherry+, and double positive cells (both nestin+ and mCherry+) in culture. H) Quantification data showing the population of ki67 positive cells, proliferation marker, in AAV-mCherry and AAV-SOX2-mCherry transduced cells. I) The percentage of confluence for AAV-SOX2-mCherry and AAV-mCherry transduced cells at any given time point acquired using IncuCyte live imaging. J) Cell cycle analysis on AAV-SOX2-mCherry and AAV-mCherry transduced astrocytes showing significant increase in S phase and G2/M phase of SOX2 transduced cells indicating the presence proliferating cells. Data represents Mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 . N=3-5 independent cultures/group. Scale bar=100 µm.
Article Snippet: The following primary antibodies (neural and astrocyte markers) were used: mouse monoclonal anti-β3-tubulin/TUJ1, (1:500, biolegend), goat monoclonal anti-Glial Fibrillary Acidic Protein (GFAP, 1:500, Abcam), chicken polyclonal anti-mCherry (mCherry, 1:1000, Abcam), rabbit monoclonal anti-NeuN (Neun, 1:1000, Abcam)/ rabbit monoclonal anti-Microtubule-associated protein 2 (MAP2, 1:1000, Abcam), rabbit polyclonal anti-ki67 (1:300, Novus), and rabbit polyclonal anti-OCT3/4 (1:100, Invitrogen).
Techniques: Transduction, Staining, Marker, Imaging, Cell Cycle Assay