RANKL Is Involved in Runx2-Triggered Hepatic Infiltration of Macrophages in Mice with NAFLD Induced by a High-Fat Diet

Figure 4

RANKL involved in Runx2-induced macrophage recruitment in vitro. mHSCs infected with lentivirus carrying control scrambled shRNA (Lenti-ctrl), lentivirus overexpressed vector (Lenti-Runx2), or shRNA specific for Runx2 (Lenti-shRunx2) for regulation of Runx2. siRNA silenced RANKL or MCP-1 in mHSCs. (a) Western blot of mHSCs infected with lentivirus to detect the protein expression of Runx2 and RANKL. (b) ELISA results on mHSCs infected with lentivirus after forty-eight hours, to detect the concentration of soluble RANKL. (c) The mRNA expressions of Runx2 and RANKL were observed via real-time PCR in mHSCs transfected with Lenti-ctrl, Lenti-Runx2, or Lenti-shRunx2. (d) mHSCs treated with lentivirus for Runx2 or siRNA for RANKL or MCP-1 and cocultured with macrophages to perform transwell assay; images were amplified 200x. (e) Quantification for migrated macrophages per field (×200). Results from 3 independent experiments are presented. vs. controls and vs. controls.

(a)

(b)

(c)

(d)

(e)