Fig 1: Trx1 was required for AET-induced improvement of cardiac function and heart remodeling. A, Results of echocardiography; B, Analyses of LVIDd, LVIDs, LVEF and FS.C, Masson’s Trichrome staining in paraffin embedded sections of heart tissue, cardiomyocytes (red) and collagen fibers (blue) were shown. D, Cardiac fibrosis in the infarcted heart was evaluated by the collagen volume fraction (CVF). E, Analysis of heart weight / body weight (HW/BW). Values were expressed as mean ± SEM, n = 6. LVIDd, left ventricular internal diameter at end diastolic; LVIDs, left ventricular internal diameter at end systolic; FS, fractional shortening; EF, ejection fraction.
Fig 2: Trx1 inhibited H2O2-induces ROS production. A and B, H9c2 cells were treated with different concentrations of TXN (0, 10 ng/µl, 20 ng/µl, 30 ng/µl, 50 ng/µl) for 24 h (A) and H2O2 (0, 0.1 mM, 0.2 mM, 0.4 mM and 0.8 mM) for 4 h (B), cell viability was determined by CCK8 assay; C, ROS level was detected by DCFH-DA staining (green) in H9C2 cells. D, The IOD of ROS in each group were analyzed in each group. Values were represented as means ± SED, Scale bar = 200 µm Trx1, Thioredoxin1; TXN, human recombinant Trx1 protein.
Fig 3: Trx1 mediated the expression of ER stress-related protein and cell apoptosis in H9c2 cells. A and B, Expression of TXNIP, GRP78, CHOP and caspase12 in H9c2 cells was examined with Western blotting; C, Cell apoptosis was detected by TUNEL staining (red) and DAPI labeled the nucleus (blue) in H9C2 cells. D, The percentages of TUNEL-positive cells in each group were analyzed. Values were represented as means ± SED, Scale bar = 200 µm. ER, endoplasmic reticulum; TXNIP, Thioredoxin Interacting Protein; TUNEL, transferase-mediated deoxyuridine triphosphate-biotin nick end labeling; DAPI, 4',6-diamidino-2-phenylindole.
Fig 4: AET up-regulated Trx1 expression and inhibited production of ROS in the infarcted heart. A-C, Protein expression of Trx1 and TXNIP in the heart following MI was examined with Western blotting; D and E, DHE staining and analysis of fluorescence IOD in each group. The ROS level was detected by DHE fluorescence examination (red). Scale bar = 200 µm; Values were expressed as mean ± SEM, n = 5. AET, aerobic exercise training, Trx1, Thioredoxin1; TXNIP, Thioredoxin Interacting Protein; ROS, reactive oxygen species; DHE, Dihydroethidium; Sham, the Sham group; MI, the myocardial infarction group; ME, MI with AET group; MEP, ME with Trx1 inhibitor PX-12 group.
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