Growth arrest specific 6 ameliorated inflammation and portal pressure through activating efferocytosis in cholestasis and portal hypertension

Figure 7 Effects of recombinant growth arrest specific 6 on macrophage efferocytosis and liver sinusoidal endothelial cells injury in vitro. A: Western blotting analysis with quantifications of MerTK phosphorylation in RAW264.7 cells treated with vehicle or recombinant growth arrest specific 6 (rGas6); B: Transcription levels of Mrc1 and Nos2 measured by qRT-PCR in RAW264.7 cells treated with vehicle or rGas6; C: Western blotting analysis with quantifications of CD206, iNOS, tumor necrosis factor α (TNF-α) and interleukin (IL)-6 in RAW264.7 cells treated with LPS or rGas6; D: Transcription levels of Mrc1, Nos2, TNF-α and IL-6 measured by qRT-PCR in RAW264.7 cells treated with LPS or rGas6; E: Efferocytosis assay of RAW264.7 cells treated with vehicle or rGas6. Apoptotic cells were labeled with CFSE; F: IF staining of efferocytosis and macrophage reparative phenotype in RAW264.7 cells treated with vehicle or rGas6; G: Quantitative analysis of efferocytosis and CD206 intensity of RAW264.7 cells treated with vehicle or rGas6; H: Diagram of the co-culture of macrophages and liver sinusoidal endothelial cells (LSECs); I: Western blotting analysis with quantifications of eNOS phosphorylation in hLSECs treated with conditioned media (CM) from macrophages; J: Transcription levels of sinusoidal marker (LYVE1) and capillary markers (VWF and CD34) measured by qRT-PCR in hLSECs treated with CM from macrophages. ^aP < 0.05, ^bP < 0.01. rGas6: Recombinant growth arrest specific 6; TNF-α: Tumor necrosis factor α; IL: Interleukin.