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. 2016 Aug 24;3:16056. doi: 10.1038/mtm.2016.56

Figure 6.

Figure 6

Expression of active Rap1a in RPE cells reduces cell barrier compromise from TNFα by modulating IQGAP1 interactions with β-catenin and IQGAP1 phosphorylation. Rap1 activity assay in (a) retinal pigment epithelial (RPE) cells exposed to PBS, H2O2 (10 µmol/l), or human recombinant TNFα (20 ng/ml) overnight and (b) and (c) coimmunoprecipitation of β-catenin and pan-cadherin and western blot of pan-cadherin (b, representative gel images and c, quantification of densitometry of b; fold change over Ad-GFP/PBS) and (d–f) coimmunoprecipitation of IQGAP1 and β-catenin and phosphorylation of IQGAP1 in RPE cells transduced with Ad-GFP or Ad-63E and treated with PBS or TNFα (20 ng/ml) for 5 hours (d, representative gel images and e and f, quantification of densitometry of d) (g) Transepithelial resistance (TER) and (h) normalized CEC transmigration (fold change over Ad-GFP/PBS) of RPE cells transduced with Ad-GFP or Ad-63E and treated with PBS or TNFα (20 ng/ml) for 24 hours (*P < 0.05, **P < 0.01 and ***P < 0.001 versus Ad-GFP/PBS; †P < 0.05, ††P < 0.01 and †††P < 0.001 versus Ad-GFP/TNFα and ‡‡‡P < 0.001 versus Ad-63E/PBS).