Fig. 2: PATJ depletion in human brain microvascular endothelial cells (hCMEC/D3) causes the loss of tight junctions and confers a dramatic morphological change.

A WB showing PATJ expression in three clones. Five main species were identified (230, 200, 135, 80, and 60 kDa). GAPDH was used as loading control. B Relative quantification of the integrated band intensities of three different WBs for PATJ species. Bars represent mean intensity ± SD. *p < 0.05; **p < 0.01; ***p < 0.005 and ****p < 0.0001 as compared to CTL cells (ANOVA followed by Bonferroni’s test). C PATJ KD3 cells lost their cell-cell interactions, as evidenced by immunofluorescence against the TJs protein zonula occludens 1 (ZO-1). D Differential expression of TJs proteins in the three PATJ KD cells. WB analyses showed that the loss of TJs in PATJ KD3 was due to the depletion of its constituent proteins ZO-1, occludin, and claudin-11. The two intermediate PATJ KD clones (KD1 and KD2) had different immunoreactive bands for occludin and ZO-1 than CTLs. E Transwell permeabilization assay evidenced a complete loss of permeability of the PATJ KD3 cells. *p < 0.05 as compared to CTLs (Student’s t-test).