Analysis of the effect of ADAM17 silencing on lymphatic endothelial cells (LEC) proliferation.

<p>(<b>A</b>) RT-PCR analysis of the expression of members of the EGF receptor family in wild type (WT), M and S1. Positive control (ctrl+)–cDNA from cells that express particular receptors. Reaction mixtures after 40 cycles of quantitative RT-PCR were subjected to electrophoresis in the presence of ethidium bromide (EtBr). The result (shown in photographic negative) is representative of 3 performed experiments. (<b>B</b>) Western blotting analysis of EGFR and HER2 in LEC lysates. (<b>C</b>) Western blotting analysis of HB-EGF in cell lysates and media of LEC sublines M, S1, S2 and of M exposed for 48 h to 25 μM GM6001 (GM). mHB-EGF, membrane HB-EGF; sHB-EGF, soluble HB-EGF. (<b>B, C</b>) β-actin was used as a loading control of lysate proteins; a fragment of blot stained with Coomassie Brilliant Blue after antigen detection procedure was used as a loading control of media proteins. Representative pictures of three independent experiments are shown. (<b>D</b>) Changes in the number of WT, M and S1 cultured in basal medium acquired by cell counting. Bars represent mean ± SD of three independent experiments performed in triplicates.