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. 2020 Apr 25;12(5):276. doi: 10.3390/toxins12050276

Figure 2.

Figure 2

Generation of MNs from iPSC line IMR90 with protocols based on Du et al. [38], Maury et al. [39], and Kroehne et al. [40]. Representative images of the differentiation progress are shown with staining of molecular markers characteristic for the differentiation stages of undifferentiated iPSCs IMR90 (top panel), pMNs, and mature MNs (bottom panel). Undifferentiated pluripotent stem cells can be identified by nuclear expression of transcription factors SRY (Sex determining region Y) box 2 (SOX2) and octamer-binding protein 4 (OCT4) and form round colonies with smooth edges. pMNs exhibit transcription factors oligodendrocyte transcription factor 2 (OLIG2) and NK6 homeobox 1 (NKX6.1) staining in the nucleus on day 12 for the protocol based on Du et al. [38], day 9 for the protocol based on Maury et al. [39], and day 6 of MN specification for the protocol based on Kroehne et al. [40]. Mature MNs express choline O-acetyltransferase (CHAT) and insulin gene enhancer protein ISL-1 and were analyzed on day 28 of the protocol based on Du et al. [38], day 32 of the protocol based on Maury et al. [39], and day 21 of the protocol based on Kroehne et al. [40]. The pan-neuronal marker β3-tubulin (TUJ1) was used for staining of neurons. Controls for the antibodies can be seen in Figure A1. Scale bar = 50 µm.