Phospho-MER/TYRO3 (Tyr753/Tyr685) Antibody - #AF8443

Fig. 5. MSC-EVs promotes macrophage efferocytosis through activation of the MerTK/ERK/COX2 signaling pathway. A BMDMs labeled with F-Actin (red) were incubated with apoptotic neutrophils labeled with PKH67 (green), and the uptake efficiency was assessed, 200–300 cells in each field were quantified, Scale bar = 50 µm. B BMDMs were incubated with MSC-EVs for indicated times, and the expression of p-MerTK, MerTK, p-AKT, AKT, p-ERK, ERK in BMDMs was detected by western blot. C BMDMs were pretreated with MSC-EVs or PBS control before efferocytosis. Then, BMDMs labeled with F-Actin (red) were incubated with apoptotic cells labeled with pHrodo (green) to assess efferocytosis, 200–300 cells in each field were quantified, Scale bar = 50 µm. D Mice were subjected to HIRI after being pretreated with MSC-EVs or PBS, representative CD68 and TUNEL staining of liver sections were performed (n = 5 per group), in situ efferocytosis was marked by white arrows, 300–500 cells in each field were quantified, Scale bar = 50 µm. E BMDMs pretreated with MSC-EVs showed altered expression levels of p-MerTK, MerTK, p-ERK,and ERK when co-incubated with apoptotic neutrophils as detected by western blot analysis. F BMDMs were pre-treated with MSC-EVs for 2 h, followed by incubation with (+) or without (-) ACs for 45 min. After an additional 6 h of incubation, COX2 protein expression was assessed in the cells using western blot analysis. (G) PGE2 production in cultural supernatant were assessed by ELISA. Data were presented as the mean ± SD.

Figure 10. Western blot analysis was applied to analyze the expression and phosphorylation of c‐Met and MERTK in HCT116 cells with staurosporine or increasing doses of compound 17c. All experiments were performed at least three times. 

Fig. 6. Paeoniflorin enhanced the expression of MerTK and inhibited MMP9 activity in ARPE-19 cells. ARPE-19 cells were cultured in standard medium or different high-glucose media (HG, 25 mM, 50 mM, or 75 mM) for 24 h (n = 3). Paeoniflorin (0.1 μM, 1 μM, or 10 μM) was used to pretreat the cells for 4 h before HG stimulation, ADAM17 inhibitor TAPI-1 (20 μM) was used 30 min before HG stimulation and the cells were then cocultured for 24 h (n = 3). (A–C) Western blot and quantification of p-MerTK, p-ADAM17, and PDI protein levels under HG conditions. (D–F) Western blot and quantification of p-MerTK, p- ADAM17, and PDI protein levels after paeoniflorin treatment. (G) Expression of p-MerTK after TAPI-1 treated in RPE. (H–I) Paeoniflorin upregulated SOCS3 expression and inhibited MMP9 activity. *P < 0.05, **P < 0.01.