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. 2023 Mar 7;16(3):dmm049750. doi: 10.1242/dmm.049750

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© 2023. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 2. — Characterization of WLC, HC and NPC spheroids. (A) Hematoxylin and Eosin (H&E) staining and CD68, αSMA, HNF4A and CK19 immunohistochemistry (IHC) of Day 7 WLC, HC and NPC spheroids for the indicated hepatic lineage markers. Scale bar: 100 µm. (B) Quantitative RT-PCR of Day 7 WLC, HC and NPC spheroids for the two indicated HC marker genes. Table below shows the unpaired two-tailed Student’s t-test comparison of the indicated culture conditions; *P<0.05, ***P<0.001, ****P<0.0001. (C) Albumin IHC and periodic acid–Schiff (PAS) staining of WLC spheroids (Sph^WLC) and HC spheroids (Sph^HC). Scale bar: 50 µm. (D) Albumin levels in the spheroid conditioned medium measured by enzyme-linked immunosorbent assay on Day 1, 4 and 7. A base level of albumin was detected in NPC spheroids (Sph^NPC) because a 0.1% bovine serum albumin solution was used to dissolve the growth factors supplemented in the spheroid culture medium. (E) Low-density lipoprotein (LDL) uptake assay of the spheroids with (+) or without (−) anti-LDL antibody (LDL-Ab). Scale bar: 50 µm.