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. Author manuscript; available in PMC: 2021 Jun 18.
Published in final edited form as: Nat Protoc. 2021 Jan 11;16(2):919–936. doi: 10.1038/s41596-020-00441-w

Fig. 4 |. Long-term culture–induced HBPO maturation.

Fig. 4 |

a, Whole-mount immunostaining of an HBPO organoid at day (D) 12. Left: blue, CDX2; green, PDX1; red, RFP; white, DAPI. Right: a magnified image of the left sample. Green, PDX1; red, CDX2; white, RFP. b, Whole-mount staining of an HBPO organoid at day 13. Green, E-cadherin (Ecad); red, RFP; blue, DAPI. c, Morphogenic change of the PROX1+ domain of the air–liquid interface culture. Red, PROX1-tdTomato. d, Whole-mount immunostaining of a long-term cultured HBPO. Blue, DAPI; green, E-cadherin; red, HNF4a. e, Whole-mount immunostaining of a long-term cultured HBPO. Blue, DAPI; green, PDX1; red, AFP. Right: lateral view. f, Histological analysis of a long-term cultured HBPO. Left: H&E staining; middle: immunostaining for albumin; right: DBA staining. Scale bars, 100 μm (a and e), 50 μm (b and d) and 200 μm (c and f).