Additional file 6: Figure S3. of Hypermethylation and loss of retinoic acid receptor responder 1 expression in human choriocarcinoma

Functional Assays using Jeg-3 cells overexpressing RARRES1. Jeg-3 cells were transfected with a RARRES1 pDest26 plasmid or a Mock control and Electric Cell-substrate Impedance sensing (ECIS, n = 6) was performed 48 h after transfection (A). The normalized mean impedance over a time period of 24 h of RARRES1 and Mock transfected cells and a medium control is presented (A). Efficient transfection was controlled by Western Blot analysis (B) using a Goat anti-Human RARRES1 polyclonal antibody (R&D Systems, Wiesbaden; 40 kDa) and a Rabbit anti-Human GAPDH polyclonal antibody (Santa Cruz, Heidelberg; 37 kDa). Additionally, proliferation of RARRES1 pDest26 and Mock transfected cells (C) (n = 3, respectively) was measured over a time period of 72 h using the CyQuant NF Cell Proliferation Kit (Thermo Fisher, Darmstadt). Migration of RARRES1 pDest26 and Mock transfected cells (n = 4, respectively) was measured over a time period of 48 h by Scratch Assay (D). The normalized proportion of the cell-free gap was calculated using ImageJ (Wayne Rasband, Nat. Institute of Health, USA). (TIFF 298 kb)