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. Author manuscript; available in PMC: 2018 Dec 28.
Published in final edited form as: Sci Transl Med. 2018 Feb 7;10(427):eaam8604. doi: 10.1126/scitranslmed.aam8604

Fig. 5. TG2 agonist TSG12 relaxes ASMCs and ameliorates pulmonary resistance.

Fig. 5.

(A) Pulmonary resistance (R) in OVA-challenged mice treated with vehicle (OVA), TB, or TSG12 (*P < 0.05 versus OVA and #P < 0.05 versus TB, n = 15 per group). (B) Pulmonary resistance (R) at baseline [control without house dust mite (HDM)] and HDM-challenged mice treated with vehicle (HDM), ISO (10 mg/kg), or TSG12 (100 ng/kg) or inactive CC (100 ng/kg). (*P < 0.05 versus HDM and #P < 0.05 versus baseline, n = 8 per group). Mch, methacholine. (C) Western blot analyses of TG2 (TG2) and β-actin in human ASMCs treated with control or specific siRNA inhibiting TG2 expression. Lower panel: Real-time cell analysis of control and TG2-deficient human ASMCs challenged with acetylcholine without treatment (ACh) or treated with TB (10,000 nM) or TSG12 (50 nM). (*P < 0.05 versus ACh and #P < 0.05 versus TB, n = 4 per group). (D) Optical magnetic twisting cytometry of human ASMCs challenged with acetylcholine without treatment (ACh) or with ISO (100 mM), TSG12 (10 and 100 nM) or, inactive CC (100 nM). (*P < 0.05 versus ACh and #P < 0.05 versus ISO, n = 4 per group). (E) Phase-contrast images and traction maps of human ASMCs (scale bars, 20.0 mm). (F) Traction force of human ASMCs treated with vehicle (control) or treated with TSG12 (100 nM), ISO (100 mM), or inactive CC (100 nM). Bars are the geometric means of the respective group. Cell traction data are converted to log scale before the statistical analyses. (*P < 0.05 versus control, n = 23 per group). (G) Real-time cell analysis of human ASMCs challenged with acetylcholine without treatment (ACh) or treated with salbutamol (SAL; 10 µM) or with a second treatment of either salbutamol (SAL/SAL; 10 μM) or TSG12 (SAL/TSG12; 10 nM) within 24 hours. (*P < 0.05 versus ACh and #P < 0.05 versus human ASMCs with β2-adrenoceptor desensitization, n = 4per group). (H) Real-time cell analysis of human ASMCs challenged with acetylcholine without treatment (ACh) or treated with TSG12 (10 nM) or with a second treatment (TSG12/TSG12; 10 nM) within 24 hours (*P < 0.05 versus ACh, n = 4 per group). All data are expressed as means ± SEM. Statistical analyses were performed by one-way ANOVA with the Games-Howell/LSD post hoc test.