Figure 2

Targeting HLX1 with siRNA abrogates its basal expression and induction by HGF. (A) DY-547-labeled control siRNA (siCtrl) was transfected into HTR-8/SVneo cells for 24 h. The transfection efficiency, as determined by flow cytometry, is presented. Black line, non-transfected (NT) cells; gray line, siCtrl cells. (B) The steady-state mRNA level of HLX1 in indicated HTR-8/SVneo cells was examined by qPCR and calculated as the ratio of HLX1 to ß-actin. (C) The protein level of HLX1 in indicated HTR-8/SVneo cells was examined by Western immunoblot. A representative gel image is presented on the left and quantifications of HLX1 to β-actin signal ratio on the right.(D) HTR-8/SVneo cells were transfected with either siCtrl or siHLX1 and treated with or without 20 ng/mL HGF. The steady-state mRNA level of HLX1 was examined by qPCR and calculated as the ratio of HLX1 to Β-actin. (E) HTR-8/SVneo cells were treated as in (D) and the protein level of HLX1 was examined by Western immunoblot. A representative gel image is presented on the left and quantifications of HLX1 to β-actin signal ratio on the right. Quantitative data are presented as mean ± SD for three independent experiments. For (B) and (C), the relative ratio in NT cells arbitrarily defined as 1. *P < 0.01, vs. NT and siCtrl cells. For (D) and (E), the relative ratio in siCtrl-transfected cells arbitrarily defined as 1. *P < 0.01, vs. siCtrl cells.