Figure 2. The Ron receptor is expressed in the murine MH-S cell line and negatively regulates TNFα production following LPS challenge.

(A) Western analysis of Ron protein levels from increasing amounts of MH-S cell lysates. Lysates from the murine prostate cancer cell line, TRAMP-C1, was used as a positive control. (B) Ron transcript levels were measured by qRT-PCR from RNA isolated from primary alveolar macrophages and MH-S cells. Similar levels of Ron were observed in both cell types. Expression was normalized to an internal control as described in the Materials and Methods section. (C) Western analysis of Ron protein levels from primary broncho-alveolar lavage cells (BAL) and MH-S cell lysates (150µg). (D, E) MH-S cells were pretreated with or without HGFL (400ng/ml) overnight followed by LPS (1µg/ml) stimulation. Cells and culture supernatants were collected 4 hours after final stimulation and assayed for TNFα transcript (D) and protein levels (E), respectively. Ligand-induced Ron signaling blocks the production of TNFα transcripts (D) and protein TNFα (E). Data are expressed as the mean ± standard error performed in triplicate. The data is representative of three independent experiments with similar results. *, p<0.05 compared to the corresponding LPS alone treated group.