Figure 1 | Scientific Reports

Figure 1

From: Inhibition of C1-Ten PTPase activity reduces insulin resistance through IRS-1 and AMPK pathways

Figure 1

15,16-Dihydrotanshinone I (DHTS) inhibits C1-Ten protein tyrosine phosphatase (PTPase) activity. (A) C1-Ten PTPase inhibitor screening. The malachite green assay was performed with 1.5 μg purified C1-Ten protein in the presence of DMSO (NT) or indicated compound (10 μM). Vanadate (NaV) was used as a positive control. Assays were repeated three times in duplicate. Data are presented as the mean ± standard error of the mean (SEM). (B) DHTS inhibits C1-Ten PTPase activity with an IC50 of 4.3 µM. Assays were repeated three times in duplicate. Data are presented as the mean ± SEM. (C) L6 myoblasts were pre-treated with 5 mM N-acetyl-L-cysteine (NAC) for 30 min, and the cells were co-treated with NAC and the indicated compound (DMSO, 10 μM DHTS, 1 mM H2O2, or 25 μM Menadione) for 1 h. NT, DMSO control. Data are presented as the mean ± SEM (n = 4); *P < 0.05, **P < 0.01, and ***P < 0.001; NS, not significant. (D) L6 myoblasts were pre-treated with 5 mM NAC for 30 min, and the cells were co-treated with NAC and the indicated compound (DMSO, 10 μM DHTS, 1 mM H2O2, or 25 μM Menadione) for 1 h. NT, DMSO control. To determine intracellular ROS generation, the cells were loaded with 5 μM CM-H2DCFDA for 30 min. After trypsinization, the fluorescence intensity was measured by flow cytometry. Data are presented as the mean ± SEM (n = 4); **P < 0.01; NS, not significant.

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