Ju Hwan Song (Korea Basic Science Institute)

Ju Yeon Lee (Korea Basic Science Institute)

Ju Hwan Song (Korea Basic Science Institute)

Hyeon-Gu Yeo (Korea Research Institute of Bioscience and Biotechnology)

Jinyoung Won (Korea Research Institute of Bioscience and Biotechnology)

Jeongwoo Hong (Meteor Biotech)

Jae-Hun Lee (Institute for Basic Science)

Kyung Seob Lim (Korea Research Institute of Bioscience and Biotechnology)

Chang-Yeop Jeon (Korea Research Institute of Bioscience and Biotechnology)

Junghyung Park (Korea Research Institute of Bioscience and Biotechnology)

Sumin Lee (Meteor Biotech)

C. Justin Lee (Institute for Basic Science)

Young Hye Kim (Korea Basic Science Institute)

Kyoung Heon Kim (Korea University)

Youngjeon Lee (Korea Research Institute of Bioscience and Biotechnology)

Ju Yeon Lee (Korea Basic Science Institute)

Reactive astrocytes are key mediators of post-stroke tissue remodeling, yet their spatially resolved molecular profiles in the primate brain remain largely unexplored. Here, we present a spatial proteomics workflow targeting astrocyte-enriched microdomains in the ischemic cortex of nonhuman primates (NHPs). Photothrombotic stroke was induced unilaterally in the motor cortex of two NHP subjects, with the contralateral hemisphere as an internal control. Reactive astrogliosis was confirmed by GFAP immunostaining, showing hypertrophic morphology along the peri-infarct border. Astrocyte-enriched regions were selectively isolated as 100 × 100 × 30 μm³ sections using Laser Microdissection (LMD) and Spatially resolved Laser Activated Cell Sorting (SLACS), yielding 320 samples (160 per subject; 80 stroke + 80 control). Peptides were prepared by one-pot digestion and analyzed by DIA LC–MS/MS on a Thermo Orbitrap Fusion Lumos with LFQ. Samples were filtered using three QC metrics (iRT peptide intensity, total precursor intensity, and precursor count), retaining 233 samples after outlier removal. Of 24,696 proteotypic precursors identified, 3,392 with missing values below 50% were selected and aggregated to 859 proteins. Key astrogliosis markers (GFAP, VIM) and neuronal integrity markers (NEFL, NEFH) were successfully detected with preserved spatial context, providing a high-depth proteomic resource for investigating region-specific astrocyte responses in the primate ischemic cortex.​