Fig. 7

Oxymatrine targets toll-like receptor 2 (TLR2) to induce magakaryocyte (MK) differentiation and thrombopoiesis. (A) Venn diagram exhibiting the shared targets of oxymatrine and thrombocytopenia. (B) The interaction of the shared targets analyzed by the Cytoscape_v3.7.1 software. (C) Protein-protein interaction (PPI) network for identifying the core target of oxymatrine against thrombocytopenia. (D) The molecular docking model for TLR2 and ligands (oxymatrine). (E) After oxymatrine (10, 20, and 40 ?M) intervened Meg-01 cells for five days, the representative immunoblot images and biochemical quantification of TLR2 were enhanced. (F) The histogram shows the relative density of TLR2 to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (n = 3 per group). (G) Cellular thermal shift assay (CETSA) analysis of TLR2 degradation damage under different temperatures. (H) The histogram shows the relative density of TLR2 to GAPDH (n = 3 per group). (I) The drug affinity responsive target stability assay (DARTS) assay indicated oxymatrine blinding to the TLR2 in a concentration-dependent manner. Treatment with protease (1:1000) was conducted for 10 min at 40 °C (n = 3 per group). (J) DARTS assay for target validation. The stability of protein TLR2 was enhanced upon the treatment of oxymatrine (100 ?M) in Meg-01 lysates. Concentration gradient (1:500, 1:1000, and 1:1500) protease was added for 60 ?g/mL stock for 10 min at 40 °C (n = 3 per group). Data represent the mean ± standard deviation (SD) of three independent experiments. ?P < 0.05, ??P < 0.01, and ???P < 0.001 vs. the control group. NF?B1: nuclear factor kappaB subunit 1; DMSO: dimethyl sulfoxide.