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. 2017 Nov 6;7:14514. doi: 10.1038/s41598-017-15108-y

Figure 4.

Figure 4

Thiostrepton effect on tight junction components of Caco-2 cell monolayers. Caco-2 cell monolayers were incubated with vehicle or 1, 10 or 100 µM thiostrepton. Triton-X (1%)-soluble and -insoluble lysates were collected and immunoblotted for claudin-1 (CLDN1), claudin-3 (CLDN3) or claudin-4 (CLDN4) (a). β-actin served as the loading control. Pre-cropped blots are included in Supplementary Figure 6. Relative protein density was calculated as the ratio of the protein density to the density in the vehicle control (b). Data are means ± SD (n = 3). *P < 0.05 vs. vehicle-treated group, as determined by using Dunnett’s test. (c) Caco-2 cells were treated with or without 100 µM thiostrepton for 24 h, and then cell lysates were subjected to qRT-PCR analysis for CLDN1, CLDN3, or CLDN4. Data are means ± SD (n = 3). *P < 0.05 vs vehicle-treated group, as determined by using Dunnett’s test. (d) Immunofluorescence localisation of CLDN1, CLDN4, occludin (OCLN) and ZO-1 after treatment with 100 µM thiostrepton or no treatment for 24 h. Bar = 10 µm.