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. 2017 Feb 7;12(2):e0170346. doi: 10.1371/journal.pone.0170346

Fig 5. Effects of p-ERK1/2 and p-p38MAPK inhibitors on occludin band-shift in CMEC/D3 cells.

Fig 5

Cells were pretreated with or without 10 μM U0126 and 10 μM SB202190 for 15 min and then stimulated with or without 10 ng/mL TNFα for 15, 30, 60 min (A and C). (B) Testing the effects of inhibitors alone. Cell lysates were collected and occludin, P- ERK, T-ERK, P-p38, T-p38 and β-actin expression pattern was analyzed by immunoblotting assay. Quantification of the proportions of occludin upper band at 15 min of TNFα and with inhibitors (blots enclosed in brackets) were determined through PIupper /PItotal/PIβ-actin and then normalized to control. Results are mean ± SD from 4 or more experiments. Two-way ANOVA revealed significant main effects of TNFα and the inhibitors and a significant interaction (p<0.0001 for each). Bonferroni post-test showed significant differences between TNFα-treated groups as compared to their respective controls (as indicated by the letter “a”), SB202190 alone as compared to control/no inhibitors (as indicated by the letter “c”) and TNFα-treated groups with vs. without the inhibitors (as indicated by the letter “b”).