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. 2020 Aug 28;8(4):1805997. doi: 10.1080/21688370.2020.1805997

Figure 3.

Figure 3.

Effects of EW-7197 and TNFα-antibody on epithelial permeability and tight junction molecules treated with HMGB1 in 2.5D Matrigel culture of HLE cells. (a) Phase-contrast images and FD-4 assay of 2.5D Matrigel culture of HLE cells pretreated with 10 μM EW-7197 (EW) 2 h before treatment with 100 ng/ml HMGB1 for 24 h. Scale bar, 20 μm. (b) Immunocytochemistry for OCLN and LSR in 2.5D Matrigel culture of HLE cells pretreated with 10 μM EW-7197 (EW) 2 h before treatment with 100 ng/ml HMGB1 for 24 h. Scale bar, 20 μm. (c) Phase-contrast image and FD-4 assay in 2.5D Matrigel culture of HLE cells pretreated with 40 μM TNFα-antibody (TNFαab) 2 h before treatment with 100 ng/ml HMGB1 for 24 h. Scale bar, 20 μm. (d) Immunocytochemistry for OCLN and LSR in 2.5D Matrigel culture of HLE cells pretreated with 40 μM TNFα-antibody (TNFαab) 2 h before treatment with 100 ng/ml HMGB1 for 24 h. Scale bar, 20 μm. (e) Western blotting for CLDN-1, −2, −4, −7, LSR, TRIC and actin in 2.5D Matrigel culture of HLE cells cells pretreated with 10 μM EW-7197 2 h before treatment with 100 ng/ml HMGB1 for 24 h. (f) Western blotting for CLDN-1, −2, −4, −7, LSR, TRIC and actin in 2.5D Matrigel culture of HLE cells pretreated with 40 μM TNFα-antibody (TNFαab) 2 h before treatment with 100 ng/ml HMGB1 for 24 h. The corresponding expression levels of E and F are shown as bar graphs. Control vs *p < .05