Journal: bioRxiv

Article Title: Border-forming wound repair astrocytes

doi: 10.1101/2023.08.25.554857

Figure Lengend Snippet: a. Violin plots of snRNAseq detected genes enriched in reactive astrocyte clusters compared with uninjured. b. Immunohistochemistry of proteins Tyrobp and Trem2 in LBA after SCI. c,d. Selected astrocyte DEGs exhibiting different patterns of expression changes in the form of acute rise followed by decline (c) or delayed but persistent increase (d) after SCI as detected by Astro-RiboTag RNAseq. e,f,g. Immunohistochemistry of proteins C3, S100a6 and Prdx6 in LBA after SCI. Box in e shows location of detail. h. Scatter plot comparing mean log 2 FC and mean FPKM of 1927 pARGs upregulated at least 2-fold from 28-70d after SCI as detected by Astro-RiboTag RNAseq. i. Scatter plot comparing mean log 2 FC and mean enrichment (log 2 FE) of 1927 pARGs upregulated from 28-70d after SCI. j. GO Biological Processes associated with 1927 pARGs upregulated at least 2-fold from 28-70d after SCI. k. Summary schematic showing local astrocyte responses to CNS tissue damage by dedifferentiation, proliferation and transcriptional reprogramming into border forming wound repair astrocytes.

Article Snippet: Primary antibodies goat anti-A2m (1:300, AF1938; R&D Systems); rabbit anti-Aldh1l1 (1:1000; Ab87117; Abcam, Cambridge, MA, USA); sheep anti-BrdU (1:800, NB-500-235; Novus); rat anti-C3 (1:400, NB200-540; Novus); goat anti-CD13 (1:600, AF2335; R&D Systems, USA); rat anti-Cd44 (1:400, 14-0441-82; Invitrogen); rat anti-CD68 (1:1000, MCA1957; Biorad, USA); rabbit anti-Cd74 (1:200, A13958; Abclonal); rabbit anti-Cdsn (1:800,13184-1-AP; Proteintech); goat anti-Cxcl10 (1:200, AF-466; Novus); rabbit anti-Dnali1 (1:500, 17601-1-AP, Proteintech); rabbit anti-Fxyd1 (1:800, A15082, Abclonal); rabbit anti-GFAP (1:2,000; GA524; Z033401-2; Dako/Agilent Tech., CA); rat anti-GFAP (1:1,000, 13-0300; Thermofisher, USA); rabbit anti-hemagglutinin (HA) (1:1000, H6908, Sigma); goat anti-Gpc5 (1:200, AF2607; R&D Systems); rabbit anti-Gpx1 (1:200, 29329-1-AP; Proteintech); goat anti-HA (1:800, NB600-362, Novus Biologicals); rabbit anti-H2-Ab1 (1:200, A18658; Abclonal); rabbit anti-Hpse (1:200,24529-1-AP; Proteintech);guinea pig anti-Iba1 (1:1000, 234004; Synaptic Systems, USA); rabbit anti-Iba-1 (1:800, 019-19741; Wako, Osaka, Japan); rabbit anti-Id3 (1:500; 9837; Cell Signaling); rabbit anti-Kcnj10 (Kir4.1) (1:400, APC-035; Alomone labs)rat anti-Lgals3 (1:200, 14-5301-82; ThermoFisher); rabbit anti-Lxn (1:500, 13056-1-AP; Proteintech); rabbit anti-Mfge8 (1:200, A12322; Abclonal); rabbit anti-Mmp12 (1:200, 22989-1-AP; Proteintech); goat anti-Myoc (1:400, AF2537; Novus); guinea pig anti-NeuN (1:1000, 266004; Synaptic Systems); rabbit anti NeuN (1:1000, ab177487, Abcam); guinea pig anti-Olig2 (1:800, ABE1024; Millipore); rabbit anti-Olig2(1:200, AB9610; Millipore); rabbit anti-Padi2 (1:300,12110-1-AP; Proteintech); rabbit anti-Prdx6 (1:500, 13585-1-AP; Proteintech); sheep anti-S100a6 (1:300, AF4584; R&D Systems); rabbit anti-S100a6 (1:200, A3461; Abclonal); goat anti-Serpina3n (1:200, AF4709; R&D Systems); goat anti-Sox9 (1:800, AF3075; R&D Systems); rabbit anti-Sox9 (1:800, 702016; ThermoFisher); goat anti-Sox10 (1:500, AF2864; R&D Systems); guinea pig anti-tdT (RFP) (1:1500,390-004; Synaptic systems); rabbit anti-RFP (1:1500,600-401-379; Rockland); rabbit anti-Timp1 (1:800, 16644-1-AP; Proteintech); sheep anti-Trem2 (1:400, AF1729; Novus); rabbit anti-Tyrobp (1:400,12492S; Cell Signaling); rat anti-Vim (1:200, MAB2105; Novus).

Techniques: Immunohistochemistry, Expressing