Fig. 4

Association between CD63, Iba1, and prognosis in glioblastomas. a Kaplan-Meier curve for glioblastoma patients stratified into three groups based on total CD63 score. b Association between WHO malignancy grade and CD63 mRNA expression levels in the TCGA. c Using the TCGA dataset, low CD63 levels were significantly associated with improved survival in glioblastoma patients when dichotomized at the 25th percentile. d-f Double-immunofluorescence staining with CD63 (green) and Iba1 (red) using DAPI as nuclear staining (blue). g Software-based classification was performed using a threshold-based algorithm. Cells were assigned color labels based on their expression of CD63 and Iba1. Cells co-expressing CD63 and Iba1 are illustrated as white-labeled nuclei, cells only expressing Iba1 are depicted as red-labeled nuclei, cells only expressing CD63 are depicted as green-labeled nuclei, while negative cells are depicted as blue-labeled nuclei. h The areas of double-positive nuclei were calculated as fractions of the total Iba1+ nuclear area and the total CD63+ nuclear area. i Kaplan-Meier curve for glioblastoma patients included in the TCGA dataset stratified into four groups based on the CD63 and Iba1 mRNA expression levels. Scale bar: 100 μm. ***: p < 0.001. Abbreviations: AA anaplastic astrocytoma; DA diffuse astrocytoma; GBM glioblastoma