MERTK Antibody - #DF7344

Figure 11. TM9SF4 regulated cytosolic pH value and cell surface expression of 2 phagocytosis-related proteins. Cytosolic pH values of (A) HCECs and (B) BMDMs were measured with pHrodo Red Indicator by flow cytometry. Shown are representative flow cytometer experiments and a data summary. (C) Whole-cell lysates of WT or KO BMDMs were collected, and cell surface proteins were isolated using surface biotinylation methods. The total and surface expressions of CD36 and MerTK were measured. ATP1A1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were the loading control for cell membrane proteins and total cellular proteins, respectively (n = 3). Means ± SEM. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001. max, maximum; SCR, scrambled control.

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.