Journal: Scientific Reports
Article Title: Histological effects of combined therapy involving scar resection, decellularized scaffolds, and human iPSC-NS/PCs transplantation in chronic complete spinal cord injury
doi: 10.1038/s41598-024-82959-7
Figure Lengend Snippet: Therapeutic effects of scar resection and dECM hydrogel (scaffold). ( a ) Experimental design and images of scar resection and dECM hydrogel (scaffold) in the chronic transected SCI model. ( b ) Heatmap of the angiogenesis-associated genes in the scar resection and scar resection + scaffold groups. Scale, 0 < − log 10|adjusted P| < 1.00. ( c ) Examination of neovascularization at the epicenter using immunohistochemical staining for CD31. Scale bar, 1000 μm. ( d ) Quantification of the CD31+ area in the 1000-μm-wide regions at the epicenter (n = 4 each). One-way ANOVA followed by the Tukey–Kramer test; N.S., not significant; *P < 0.05. Data are expressed as mean ± SEM.
Article Snippet: The sections were then processed for histological analysis using the following primary antibodies: anti-HNA (MAB4383, mouse IgG1, 1:100, Millipore, Inc.), anti-GFAP (16825-1-AP, rabbit IgG, 1:500, Proteintech, Inc.; mouse IgG2a, Thermo Fisher, Inc.), anti-CS56 (C8035, rabbit IgG, 1:200, Sigma–Aldrich), anti-Arg1 (ab60176, goat IgG, 1:400, Abcam, Inc.), anti-Iba1 (019-19741, rabbit IgG, 1:1000, Wako), anti-CD31 (AF3628, goat IgG, 1:100, R&D Systems, Inc.), anti-ELAVL3/4 (Hu C/D) (A21271, mouse IgG2b, 1:500, Molecular Probes, Inc.), anti-APC (ab16794, mouse IgG2b, 1:300, Abcam, Inc.), STEM121 (Y40420, mouse IgG1, 1:200, Takara Bio, Inc.), anti-NF-H (rodent-specific) (ab8135, rabbit IgG, 1:500, Abcam, Inc.), and anti-βIII-tubulin (mouse IgG2b, 1:500, Sigma, Inc.).
Techniques: Immunohistochemical staining, Staining
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![a – c Mice were fed with a standard or CDAA diet for 10 weeks. a Images show Picrosirius Red staining on the liver sections. LSECs were isolated from healthy and fibrotic liver tissues. Scale bar = 100 µm. b The dot plot shows the number of LSECs isolated from fibrotic mouse liver. [mean ± SD, n = 4 mice]. c Quantitative PCR analyses to compare the expression level of various sinusoidal genes in LSECs isolated from either healthy or fibrotic mouse liver tissues. [mean ± SD, n = 4 mice]. * P < 0.05 (Mann–Whitney test). d – f Liver tissues collected from healthy pigs were processed to isolate LSECs. d FACS plot showing gating strategy to isolate <t>CD31+</t> LSECs and CD31− cells. e The dot plot shows the number of LSECs isolated from pig liver tissues. [mean ± SD, n = 4 pigs]. f Quantitative PCR analyses to compare expression of various vascular genes between CD31+ LSECs and CD31− cells. [mean ± SD, n = 4 pigs]. * P < 0.05 (Mann–Whitney test).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1496/pmc11711496/pmc11711496__42003_2025_7458_Fig3_HTML.jpg)
![a – d LSECs were isolated using perfusion independent digestion method, followed by positive selection using CD146-magnetic beads. After that, enriched LSECs were placed in culture. a Primary LSEC cultures were stained for an endothelial cell marker (ERG, in green) and nuclear marker (DAPI, in gray). On the left, images show IF-stained LSEC cultures. Scale bar = 25 µm. The yellow arrow highlights an ERG-negative cell. On the right, the dot plot shows the percentage of ERG-positive cells of DAPI-positive cells. [mean ± SD, n = 3 wells]. b Images show IF-stained LSECs with CD31 (in gray), Stabilin-2 (in green), and DAPI (in blue). Scale bar = 25 µm. c Images show IF-stained LSECs with CD32b (in gray) and ERG (in green). Scale bar = 25 µm. d Representative scanning electron microscopy images of cultured LSECs. Scale bar = 2 µm.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1496/pmc11711496/pmc11711496__42003_2025_7458_Fig4_HTML.jpg)
