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. 2023 Nov 20;13(19):e4840. doi: 10.21769/BioProtoc.4840

Figure 5. Differentiation assays.

Figure 5.

Human Schwann cells (hSCs) were stimulated and analyzed per Protocol 3C and stained with antibodies per Table 1. Low (A–B) and high magnification (C–J) images of control (non-stimulated cells in D1) and cAMP-differentiated hSCs (cells treated with 250 μM CPT-cAMP in D1 for 7 days) are shown. Notice the phenotypic conversion of the hSCs from an elongated, roughly bipolar shape to an enlarged, flattened morphology in the CPT-cAMP condition. CPT-cAMP treatment does not change S100B (A–D) and NGFR (C–D, I–J) expression. However, a proportion of the CPT-cAMP-treated hSCs express higher levels of Krox20 (G–H), periaxin (PRX, E–F), and O4 (E–H) along with lower levels of nuclear cJun (I–J), as denoted by the white arrowheads. Heterogeneity in the expression levels of myelin markers is expected (H–F). The S100B negative cells highlighted by the black arrowheads in panels A–B are fibroblasts. Nuclei were stained with DAPI (blue, C–F). The PRX antibody was courtesy of Peter Brophy. The Krox20 antibody was courtesy of Dies Meijer.