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. Author manuscript; available in PMC: 2020 Jun 6.
Published in final edited form as: Nat Protoc. 2019 Nov 15;14(12):3303–3332. doi: 10.1038/s41596-019-0220-0

Fig. 1 |. Overview of PSC-derived alveolosphere differentiation protocol.

Fig. 1 |

a, TALENs targeting strategy and edited NKX2–1GFP and SFTPCtdTomato loci after Cre-mediated antibiotic cassette excision. a adapted with permission from ref. 3, Elsevier. b, Schematic of differentiation protocol with arrows representing steps involving passaging or sorting. Markers for intermediate cell types are noted where relevant. Media components and concentrations are listed for each respective part of the protocol. c, Schematic of options for post day 30 culture of alveolospheres with recommended sort markers for days 14, 15 and 30+ cells. 3A, 3′ acceptor site; AFE, anterior foregut endoderm; ATG, ATG translation start site; DE, definitive endoderm; E, exon; hPSC, human pluripotent stem cell; pA, poly A; pPGK, phosphoglycerate kinase I promoter; puroTK, puromycin and thymidine kinase cassette; PuroR, puromycin resistance gene; SA, splice acceptor; TALEN, transcription activator-like effector nuclease.