Fig. 3 Biological function of TG2 and the interaction with MT-2.

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Fig. 3 Biological function of TG2 and the interaction with MT-2. Biological function of TG2 and the interaction with MT-2. (A) Collagen gel contraction assays of rat ASMCs at baseline (control without treatment) or treated with TB (10,000 nM) or MT-2 (60 nM) (*P < 0.05 versus baseline and #P < 0.05 versus TB, n = 3 per group). (B) Comparison of control and TG2-deficient rat ASMCs challenged with acetylcholine without treatment (ACh) or treated with TB (10,000 nM) or MT-2 (60 nM) (*P < 0.05 versus ACh and #P < 0.05 versus TB, n = 4 per group). (C) Western blot analyses of TG2 and β-actin in lung tissue from WT or TG2 knockout (TG2-KO) mice. Lower panel: Mean density of TG2 protein normalized to β-actin protein (*P < 0.05 versus TG2-KO by t test, n = 3 per group). (D) Pulmonary resistance (R) in WT and TG2-KO mice sensitized to OVA and challenged with vehicle (control) or OVA. (*P < 0.05 versus control and #P < 0.05 versus WT mice challenged with OVA, n = 8 per group). (E) Western blot analyses of MT-2 on the phosphorylation and total protein of ezrin, myosin phosphatase target subunit 1 (MYPT1) in rat ASMCs. Right panel: Corresponding densitometric analyses showing phosphorylated/total protein (%) treated with vehicle (control) or MT-2 (30 nM) (*P < 0.05 versus control, t test; n = 3 per group). (F) Western blots of TG2 and ezrin in total lysates or immunoprecipitates with TG2 antibody (IP: TG2) or control immunoglobulin G (IgG) of lung tissue from WT or TG2-KO mice. (G) Western blots of TG2 and ezrin in total lysates or immunoprecipitates with ezrin antibody (IP: Ezrin) or control IgG of lung tissue from WT or TG2-KO mice. (H) Kinetic analysis of ezrin by Biacore surface plasmon resonance with TG2 immobilized on the CM5 sensor chip. The response signals were recorded over the indicated time periods. All data are expressed as means ± SEM. Statistical analyses were performed by one-way ANOVA with the Games-Howell/LSD post hoc test. Lei-Miao Yin et al., Sci Transl Med 2018;10:eaam8604 Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works