Figure 5. The Ron receptor negatively regulates Adam17 levels in MH-S cells.

(A) Ron protein levels were measured by western analysis of cell lysates from MH-S cells, MH-S cells infected with an shRNA construct to knockdown Ron expression (Ron shRNA) or a control shRNA construct (nonsense shRNA). Each lane represents a separate independent experimental sample. Actin expression is shown as a loading control. These studies demonstrate knockdown of Ron expression in Ron shRNA infected cells. (B) Adam17 protein levels were measured by Western analysis from MH-S cells following infection with a Ron-specific shRNA (Ron shRNA) or with a nonsense shRNA control (nonsense shRNA) construct. Actin expression is shown as a loading control. Of note, following Ron knockdown in MH-S cells, the levels of Adam17 are increased. (C) Adam17 transcript levels were measured by qRT-PCR from RNA isolated from MH-S cells pretreated with HGFL (400ng/ml) or media alone, followed by stimulation with (+) or without (-) LPS (1 µg/ml). (D) Adam17 transcript levels were measured by qRT-PCR from RNA isolated from MH-S cells treated with HGFL (100ng/ml) or media alone. Expression was normalized to an internal control as described in the Materials and Methods section. In response to LPS, Adam17 RNA levels are increased and this induction can be significantly blunted by HGFL pretreatment. Data is expressed as the mean ± the standard error and is representative of several independent experiments.