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. 2018 Feb 5;128(3):1010–1025. doi: 10.1172/JCI95802

Figure 4. YAP/TAZ directly activate Irs2 transcription through TEAD binding.

Figure 4

(A) Immunoblot analyses of insulin signaling molecules in the livers of 3-month-old mice. (B) qPCR analysis of Irs2 mRNA levels in the same livers as in A. (C–G) Representative immunoblot analyses of AML12 cells infected with lentiviruses encoding shPten and/or shSav1 (C)and their insulin-induced AKT activity (D). AML12 cells were infected with a retrovirus encoding IRS2 or a control (CTL) (E), and shPten- and shSav1-expressing AML12 cells in C were transfected with siIrs2 or control (siCtl) (F). AML12 cells were infected with retrovirus encoding TAZ4SA or TAZ4SA/S51A (G). In D and F, the cells were deprived of serum for 16 hours and then treated with insulin (100 nM) for the indicated durations. (H and I) Luciferase reporter assay (I) for 293T cells expressing TAZ4SA or TAZ4SA/S51A as well as luciferase (Luc) reporter constructs that include regions of the Irs2 distal promoter and first intron (H) containing WT or deleted (TBSsΔ). Ex, exon. (J) ChIP-qPCR analysis of the binding of TAZ to the Irs2 promoter analyzed in I. AML12 cells infected with control or TAZ4SA retroviruses and subjected to immunoprecipitation with antibodies recognizing TAZ or IgG. Quantitative data in B, I, and J represent the mean ± SEM. **P < 0.01 and ***P < 0.001 versus the corresponding CTL; †P < 0.05, ††P < 0.01, and †††P < 0.001 for the indicated comparisons. One-way ANOVA (B) and Student’s t test (I and J). (A and B) n = 3 for each group. (C–J) n = 3 independent experiments; (I) n = 5 independent experiments.