Figure 8.
Reporter gene luciferase assay shows PDIA1 acting on TRβ gene regulation. Full length TRβ wt, C294A, and C298A, full length PDIA1 and Responsive Element (RE) F2, were transfected in 293T cells (in the presence and absence of T3), then luciferase activity was measured. All the TRs activation values were normalized by Renilla Luciferase activation. (A) First, T3 hormone addition in all cases increased the transactivation, this shows that in terms of gene expression, both TRβ mutants were activated in a similar way. (B) PDIA1 presence in the TRβ mutant transactivation assay led to increase TRβ-C294A and TRβ-C298A basal activation by about 3- to 4-fold. On the other hand, in the presence of T3, PDIA1 promoted minor further reduction in TRβ-C294A and TRβ-C298A activation in comparison with wt TRβ (86-fold for C294A, 89-fold for C298A, and 115-fold for TRβ wt). Statistical Analysis with One-way or Two-way ANOVA *p < 0.05, **p < 0.01.