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A , experimental design. Inflammatory demyelination of experimental autoimmune encephalomyelitis (EAE) was induced in adult mice via MOG immunization (CFA as control) at day 0 (D0), with pertussis administered i.p. on D0 and D2. RNA-seq was performed on whole spinal cords ( B ) and MACS-purified glial populations ( C-F ) at D30-D35. OLs, oligodendrocytes; Astro, astrocytes; MG, microglia. B , volcano plot of differentially expressed genes (DEGs) in spinal cord of MOG/EAE (n=4) vs. CFA mice (n=3) at D35. Top10 DEGs are labeled with gene symbols and FPKM values. C , schematic of magnetic beads-assisted cell sorting (MACS) of spinal cord glia at D30. OL, Astro, and myeloid cells (including MG) were isolated using <t>anti-O4,</t> anti-ACSA2, and anti-CD11b magnetic beads, respectively. D , purity validation of sorted population using representative markers Plp1 (OL), Tmem119 (MG), and Aldh1l1 (Astro). Expression is shown as TPM (transcripts per million). E , volcano plot of DEGs in MOG_OL versus CFA_OL. Serpina3n is highlighted in red. F , TPM expression of Serpina3n , Ctsd , Lgals3 , and Gpnmb in OLs, Astro, and MG from MOG/EAE and CFA mice. Serpina3n is specifically upregulated in OL, while Ctsd , Lgals3 , and Gpnmb are enriched and upregulated in MG.
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A , experimental design. Inflammatory demyelination of experimental autoimmune encephalomyelitis (EAE) was induced in adult mice via MOG immunization (CFA as control) at day 0 (D0), with pertussis administered i.p. on D0 and D2. RNA-seq was performed on whole spinal cords ( B ) and MACS-purified glial populations ( C-F ) at D30-D35. OLs, oligodendrocytes; Astro, astrocytes; MG, microglia. B , volcano plot of differentially expressed genes (DEGs) in spinal cord of MOG/EAE (n=4) vs. CFA mice (n=3) at D35. Top10 DEGs are labeled with gene symbols and FPKM values. C , schematic of magnetic beads-assisted cell sorting (MACS) of spinal cord glia at D30. OL, Astro, and myeloid cells (including MG) were isolated using <t>anti-O4,</t> anti-ACSA2, and anti-CD11b magnetic beads, respectively. D , purity validation of sorted population using representative markers Plp1 (OL), Tmem119 (MG), and Aldh1l1 (Astro). Expression is shown as TPM (transcripts per million). E , volcano plot of DEGs in MOG_OL versus CFA_OL. Serpina3n is highlighted in red. F , TPM expression of Serpina3n , Ctsd , Lgals3 , and Gpnmb in OLs, Astro, and MG from MOG/EAE and CFA mice. Serpina3n is specifically upregulated in OL, while Ctsd , Lgals3 , and Gpnmb are enriched and upregulated in MG.
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A , experimental design. Inflammatory demyelination of experimental autoimmune encephalomyelitis (EAE) was induced in adult mice via MOG immunization (CFA as control) at day 0 (D0), with pertussis administered i.p. on D0 and D2. RNA-seq was performed on whole spinal cords ( B ) and MACS-purified glial populations ( C-F ) at D30-D35. OLs, oligodendrocytes; Astro, astrocytes; MG, microglia. B , volcano plot of differentially expressed genes (DEGs) in spinal cord of MOG/EAE (n=4) vs. CFA mice (n=3) at D35. Top10 DEGs are labeled with gene symbols and FPKM values. C , schematic of magnetic beads-assisted cell sorting (MACS) of spinal cord glia at D30. OL, Astro, and myeloid cells (including MG) were isolated using anti-O4, anti-ACSA2, and anti-CD11b magnetic beads, respectively. D , purity validation of sorted population using representative markers Plp1 (OL), Tmem119 (MG), and Aldh1l1 (Astro). Expression is shown as TPM (transcripts per million). E , volcano plot of DEGs in MOG_OL versus CFA_OL. Serpina3n is highlighted in red. F , TPM expression of Serpina3n , Ctsd , Lgals3 , and Gpnmb in OLs, Astro, and MG from MOG/EAE and CFA mice. Serpina3n is specifically upregulated in OL, while Ctsd , Lgals3 , and Gpnmb are enriched and upregulated in MG.

Journal: bioRxiv

Article Title: Injury-transduced SerpinOLs modulate neuroinflammation and glial activation in the diseased and non-diseased CNS

doi: 10.64898/2026.01.29.702669

Figure Lengend Snippet: A , experimental design. Inflammatory demyelination of experimental autoimmune encephalomyelitis (EAE) was induced in adult mice via MOG immunization (CFA as control) at day 0 (D0), with pertussis administered i.p. on D0 and D2. RNA-seq was performed on whole spinal cords ( B ) and MACS-purified glial populations ( C-F ) at D30-D35. OLs, oligodendrocytes; Astro, astrocytes; MG, microglia. B , volcano plot of differentially expressed genes (DEGs) in spinal cord of MOG/EAE (n=4) vs. CFA mice (n=3) at D35. Top10 DEGs are labeled with gene symbols and FPKM values. C , schematic of magnetic beads-assisted cell sorting (MACS) of spinal cord glia at D30. OL, Astro, and myeloid cells (including MG) were isolated using anti-O4, anti-ACSA2, and anti-CD11b magnetic beads, respectively. D , purity validation of sorted population using representative markers Plp1 (OL), Tmem119 (MG), and Aldh1l1 (Astro). Expression is shown as TPM (transcripts per million). E , volcano plot of DEGs in MOG_OL versus CFA_OL. Serpina3n is highlighted in red. F , TPM expression of Serpina3n , Ctsd , Lgals3 , and Gpnmb in OLs, Astro, and MG from MOG/EAE and CFA mice. Serpina3n is specifically upregulated in OL, while Ctsd , Lgals3 , and Gpnmb are enriched and upregulated in MG.

Article Snippet: After centrifugation, the supernatant was gently removed, and the brain homogenate pellet was incubated separately with anti-CD11b MicroBeads (Cat# 130-093-634, Miltenyi Biotec, Germany) and anti-O4 MicroBeads (Cat# 130-094-543, Miltenyi Biotec, Germany).

Techniques: Control, RNA Sequencing, Purification, Labeling, Magnetic Beads, FACS, Isolation, Biomarker Discovery, Expressing