Fig. 3: iASPP mutants trigger heat shock response and lose their ability to regulate NF-ĸB.
From: Cardiocutaneous syndrome is caused by aggregation of iASPP mutants
A Luciferase reporter assay for detection of heat shock factor (HSF) activity using an artificial HSE promotor. U2OS cells were transiently transfected with the luciferase reporter plasmid, along with either an empty vector control, iASPP WT or a mutant. Measured activity was normalized to the empty vector control. (Mean ± SD. n = 3, ordinary one-way ANOVA, n.s. P > 0.05; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001). B Co-immunoprecipitation of endogenous HSP70 and Myc-tagged iASPP WT or mutants transiently transfected in H1299 cells. HSP70 was immunoprecipitated with an anti-HSP70 antibody and interaction with Myc-iASPP was detected using an anti-Myc antibody. n = 3. C Luciferase reporter assay for detection of p65 activity using an NF-ĸB promotor. U2OS cells were transiently transfected with the luciferase reporter plasmid, along with either an empty vector control, iASPP WT or a mutant. 24 h after transfection, cells were treated with 50 ng/ml TNFα for 6 h. Measured activity was normalized to the untreated empty vector control. (Mean ± SD. n = 3, ordinary one-way ANOVA, n.s. P > 0.05; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001).
