Kl001

Trophoblast cells derived from a 6–12 weeks-old human placenta (HTR-8 SVNeo, referred to as HTR-8 cells, ATCC #CRL-3271) were cultured in RPMI 1640 Medium (Gibco, #11875093) with 10% FBS (Sigma-Aldrich, #F4135) and 1% Penicillin–Streptomycin (Sigma-Aldrich Cat# P4333). HTR-8 cells were plated at 0.5 million cells/ml per well in a 24-well wound healing assay (Abcam, #ab242285). Twenty-four h after plating, the HTR-8 cells formed a monolayer and the inserts were removed, where after the cells were treated with vehicle (DMSO 1/500 or 1/5000 dilution, no difference in HTR-8 cell migration in response to DMSO concentration was found and data were pooled), KL001 (Tocris Bioscience, # 46-851-0), PF670462 (Tocris Bioscience™ Cat# 33-161-0), SR9009 (Tocris Bioscience, # 5855), SR8278 (Tocris Bioscience, # 4463), or KL101 (Sigma Aldrich, #SML3014) at 1 μM and 10 μM. Bright field image acquisition of the wound healing area was done using a light microscope (Leica DMi1) 0, 24 and 48 h after treatment. Data were analyzed using ImageJ/Fiji^® version 1.53 (NIH).