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Fig. 6 | Acta Neuropathologica Communications

Fig. 6

From: High-resolution transcriptomics informs glial pathology in human temporal lobe epilepsy

Fig. 6

Glial proliferation in human temporal lobe epilepsy. (a-b) Representative immunofluorescence images from the subset of TLE samples with high proliferation, as assessed by Ki67 labeling. A large portion of Ki67-positive cells co-express the microglial marker IBA1 (arrows), although Ki67 + IBA1– cells are not uncommonly seen as well (arrowheads) (a). Representative immunofluorescence images of GFAP, OLIG2 and Ki67 co-expression in TLE tissue (b). (c) Quantification of cell type-specific contribution to proliferation in the subset of TLE samples with high proliferation index, assessed by co-expression of Ki67 with OLIG2 and/or GFAP. Ki67 + cells were examined within the entire TLE tissue section, 14–58 40X fields overall. (d) Violin plot of log-normalized EGFR expression across the six TLE glial subpopulations from Fig. 4b. (e) FACS plots showing isolation strategy to purify EGFR + (CD34–CD45–) cells from fresh TLE tissue. A small but distinct population of EGFR + (DAPI–CD34–CD45–) cells are isolated, and subjected to primary cell culture along with EGFR– cells. FACS excludes dead cells (DAPI +), endothelial (CD34 +) and inflammatory (CD45 +) cells from the culture analysis. Numbers in boxes indicate % from total DAPI– cells. (f) EGFR + cells form proliferative clusters in vitro, significantly more compared to their EGFR– counterpart, when grown under low-adherence neural stem cell condition medium (n = 5 TLE samples, 10cells/µl (2000 cells/well); 1–3 wells per sample, week 2). Box-plots represent median, minimum and maximum value. P-value calculated using one-tailed U-Mann Whitney non-parametric test. (g) Representative microscope images (10X) of proliferative clusters growing from TLE EGFR + cells. (h) Human TLE tissue immunofluorescence demonstrates occasional co-localization of the rare EGFR + cells with the proliferative marker Ki67. In (a–b) nuclei are counterstained with DAPI. Scale bar = 50 µM unless otherwise specified

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