Axio vert a1

The cell area was analyzed for 30 cells for each condition after 24 h of culture using ImageJ (version 1.53e). For cytoskeleton analysis, hUC-MSCs were plated at an initial cell density of 1.5 × 10⁴ cells/well onto the glass coverslips, which were clean (control) or covered by different PEM films. The cells were cultured for 24 h and fixed in 3.7% formaldehyde, solubilized with 0.1% Triton X-100, immunostained with mouse monoclonal anti-human vinculin IgG (Merck, Darmstadt, Germany) and Alexa Fluor 488-conjugated goat anti-mouse IgG-clone A11001 (Merck, Darmstadt, Germany), and counterstained with TRITC-phalloidin (Merck, Germany). Nuclei were stained with DAPI (Merck, Germany). The specimens were mounted onto coverslips with poly(vinyl alcohol) (Dako Fluorescent Mounting Medium, Agilent Technologies, Santa Clara, CA, USA), visualized under a fluorescence inverted microscope (Axio Vert.A1, Zeiss, Dresden, Germany), and analyzed using ZEN 2.3 (Zeiss) software.

PaCa44 and MCF7 cells were fixed in 4% paraformaldehyde for 8 min and washed 3 times with PBS for 5 min each. To saturate unspecific binding sites, the cells were incubated for 45 min at RT with a blocking solution containing 5% BSA in PBS. Samples were then incubated overnight at 4 °C with anti-LHR (1:200; Thermo Fisher #8G9A2, Waltham, MA, USA) primary antibody diluted in blocking solution. After 3 washes with PBS of 10 min each, cells were incubated for 1 h at RT in the dark with specific secondary antibody (1 µg/mL) conjugated with Alexa Fluor-488 (Molecular Probes, Eugene, OR, USA). Samples were mounted in anti-bleaching medium (Dako Fluorescent Mounting Medium, Agilent Technologies, Santa Clara, CA, USA) and examined by fluorescence microscope (Axio Vert. A1, Zeiss).