Fig. 4: Effect of DSP and PPL variants on RhoA activity.
From: Desmoplakin and periplakin genetically and functionally contribute to eosinophilic esophagitis
a Active RhoA assays performed in EPC2 cells transduced with constructs encoding non-variant and mutated DSP or PPL. Statistics (versus non-variants): p.G46D, P = 0.132; p.R808C, P = 0.0691; p.Y895C, P = 0.0453; p.1067_1068del, P = 0.1013; p.N1215S, P = 0.2131; p.R1340C, P = 0.048; p.E1723Q, P = 0.0619; p.R108C, P = 0.4849; p.E632K, P = 0.0443; p.K1051V, P = 0.1589; p.L1154V, P = 0.0419; p.E1163K, P = 0.0035; p.V1377E, P = 0.8738. b Wound healing assays performed in EPC2 cells transduced with constructs encoding non-variant and mutated DSP (p.Y895C) treated with the Rho activator CN01 or the Rho kinase inhibitor Y27632. Quantification of the wound closure after 12 h was shown. Scale bar: 500 μM. Statistics: Non-variant vs. Variant, P = 0.0001; Non-variant vs. Variant + CN01, P = 0.7658; Non-variant vs. Variant + Y27632, P < 0.0001; Variant vs. Variant + CN01, P = 0.0002; Variant vs. Variant + Y27632, P = 0.0394; Variant + CN01 vs. Variant + Y27632, P < 0.0001. c Enzyme-linked immunosorbent assay of the level of active RhoA (RhoA-GTP) in protein lysates of biopsies from control individuals, non-familial patients with inactive EoE and non-familial patients with active EoE (Control, n = 3; Inactive EoE; n = 3; Active EoE, n = 5). Western blot analysis shows the expression of total RhoA in protein lysates of biopsies from each subject. Statistics: control vs. inactive EoE, P > 0.8803; control vs. active EoE, P = 0.0398; inactive EoE vs. active EoE, P = 0.0479. For panels a, b, data are representative of three experiments performed in duplicate and are presented as mean ± SEM. For panel c, data are presented as mean ± SEM, with markers representing biologically independent subjects. For panels a–c, two-tailed P-values were determined by the following tests: the one-way ANOVA test followed by a Dunnett’s multiple-comparison test (a) or Tukey’s multiple comparisons test (b, c). *P < 0.05, **P < 0.01, and ***P < 0.001. DSP desmoplakin, PPL periplakin, MW molecular weight, SEM standard error of the mean.
