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. 2021 Nov 8;9:738081. doi: 10.3389/fbioe.2021.738081

FIGURE 2.

FIGURE 2

Co-culture of HepLPCs and HUVECs promoted HepLPCs maturation. (A) VEGFA expression was measured by qRT-PCR. Data are normalized to HepLPCs. (B) Western blot analysis of the expression of CD31 (n = 3 per group). (C) The expression of ALB and CD31 in HepLPCs and vHepLPCs was determined by co-immunofluorescent staining. Nuclei were counterstained with DAPI. (D) Immunofluorescent staining analyses of the expression of mature hepatic markers after co-culture with HUVECs, ALB, AAT, CYP3A4, HNF4A and MRP. Nuclei were counterstained with DAPI. (E) Western blot analysis of the expression of ALB and CYP3A4 in two groups (n = 3 per group). (F) Quantitative analysis of ALB secretion in supernatants. Data are normalized to HepLPCs (n = 3 per group). (G) Time-lapse imaging of microvilli network in HepLPCs and vHepLPCs at 5 min to 5 h of incubation with CDFDA (1 μM). Fluorescence images show both cell types can transport CDFDA, but vHepLPCs exhibited faster transport efficiency than HepLPCs. Data are presented as means ± SEM. *p < 0.05, **p < 0.01. Scale bars, 100 µm.