Fig 2: The effects of Aroclor1254 on the barrier function of SC epithelium after inhibiting the caveolin-dependent endocytosis. The SCs were treated with or without Aroclor1254 in the presence or absence of CO as the regimen depicted in Supplementary Figure S2a. (a and b) The barrier function of SC epithelium measured by recording TER (a) or calculating the permeability of Na-F (b) across the cell monolayer. The Na-F permeability in the vehicle control on day 3 was arbitrarily set at 100%. CO treatment was found to partially resist the disruption of BTB by Aroclor1254. Each bar refers to mean±S.D. of n=3 experiments using different batches of SCs. **P<0.01, compared with the vehicle control group. ◊P<0.05; ◊◊P<0.01, compared with the CO group. #P<0.05; ##P<0.01, compared with the Aroclor1254 group. (c) Immunoblot analysis of endocytosed JAM-A, N-cadherin, and β-catenin in SCs at different time points (0, 10, 30, 60 min) after cell surface biotinylation for 30 min in the presence of Aroclor1254 (or the mixture of Aroclor1254 and CO). Total biotinylated proteins at 0 min without stripping served as the positive control. Cell lysates without pull-down were analyzed by immunoblot to confirm identical levels of the tested protein between groups with GAPDH as the loading control. (d) Line and scatter graphs summarizing the result shown in c by calculating the percentage of endocytosed protein at each data point versus the total biotinylated protein. The endocytosis of JAM-A, N-cadherin, and β-catenin showed no difference between the CO-treated cells and CO+Aroclor1254-treated cells. *P<0.05; **P<0.01, compared with the vehicle control group. ##P<0.01, compared with the Aroclor1254 group. (e) Immunofluorescence assay to observe the distribution of N-cadherin and β-catenin at SC–SC junctions on day 6 after Aroclor1254 treatment for 24 h in the presence or absence of CO, using Alexa Fluor 488-conjugated secondary antibody (green) with cell nuclei stained with DAPI (blue). Scale bar=25 μm, which applied to all micrographs in (e)

Fig 3: Re-establishment of BTB in the recipient testes after repaired SSC transplantation. A, B Distribution of N-cadherin and JAM-A (all in green) were found to be different in different tubules, depending on the status of spermatogenesis and the differentiation status of repaired SSCs. Abnormal distribution of these proteins in empty tubules devoid of germ cells (a and g) was obviously noted and gradually returned to normalcy as showed in normal wild type control testis (e and k) following the appearance of spermatocytes (b and h), round spermatids (c and i), and elongated spermatids (d and j). Spermatocytes were labeled by anti-SCP3 antibody (red), and round spermatids and elongated spermatids were identified by the corresponding nucleus shape (blue). There were no germ cells in panels a, a’, f, g, g’, and l. The boxed areas in a–d and g–j are magnified and shown in a’–d’ and g’–j’. Scale bar = 50 µm in a–l and 10 µm in a’–d’ and g’–j’

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.

Fig 5: Junctional adhesion molecule‐A (JAM‐A) expression in lung adenocarcinoma, including its histological subtypes. Upper panel, H&E staining; middle and lower panels, immunohistochemistry of JAM‐A. Lower panel (×200) is a magnified view of the rectangular area in the middle panel image (×100) for each histology.