Fig 2: Immunoflurescent and qRT-PCR analysis of occludin and claudin-5 expression in liver sinusoidal endothelial cells.(A, C) Immunofluorescent co-staining of rat liver cryosections with anti-Occludin (A, green), anti-Claudin-5 (C, green), and anti-LYVE-1 (A, C, blue) antibodies. (E) Immunofluorescent staining of a liver sample obtained from the patient 2 with anti-Occludin (green) antibody; BD – bile ducts, S – liver sinusoids. (F, G) Immunofluorescent co-staining of liver samples obtained from the patients 6 (F) and 4 (G) with anti-VE-cadherin (F, G, green), anti-CD32b (F, G, red), and anti-Occludin (F, G, blue) antibodies. Images were acquired using laser scanning confocal microscopy. Bars 11.9 µm (A, C), 47.62 µm (E, F), 14.14 µm (G). (B, D) Quantitative reverse transcriptase-PCR with mRNA isolated from rat LSECs and rat LMECs (n indicates the number of samples analyzed, error bars represent SEM). Primers specific for Occludin (B), Claudin-5 (D), and ß-Actin as normalizer were used.

Fig 3: VE-cadherin is expressed in liver sinusoidal endothelial cells in rats and humans.(A) Immunofluorescent co-staining of human liver cryosections with anti-VE-cadherin (green) and anti-CD32b (red) antibodies. (B) Immunofluorescent co-staining of rat liver cryosections with anti-VE-cadherin (green) and anti-LYVE-1 (red) antibodies. (C) Immunofluorescent co-staining of isolated rat LSECs with anti-VE-cadherin (green) and anti-Stabilin-2 (red) antibodies. Toto3 (blue) was used to counterstain the cell nuclei. Images were acquired using laser scanning confocal microscopy. Bars 11.9 µm (A, B), 14.14 µm (C). (D) Reverse transcriptase-PCR with mRNA isolated from rat hepatoma McA-RH7777 cell line (1), freshly isolated rat LMECs (2), and freshly isolated rat LSECs (3). Primers specific for VE-cadherin or ß-actin were used.

Fig 4: Expression of JAM-family members in liver sinusoidal endothelial cells.(A) Immunofluorescent co-staining of rat liver cryosections with anti-JAM-A (green), anti-VE-cadherin (red), and anti-Stabilin-2 (blue) antibodies. (B) Immunofluorescent co-staining of human liver cryosections with anti-JAM-A (green), anti-CD32b (red), and anti-Stabilin-2 (blue) antibodies. Images were acquired using laser scanning confocal microscopy. Bars 11.9 µm. (C) Quantitative reverse transcriptase-PCR with mRNA isolated from rat LSECs and rat LMECs (n indicates the number of samples analyzed, error bars represent SEM). Primers specific for JAM-A, JAM-B, JAM-C, and ß-Actin as normalizer were used.