Figure 3.

Down‐regulation of ECM remodelling gene expression in VIP‐deficient trophoblast cell lines. BeWo cell line at 60% of confluence was transfected with a VIP‐siRNA (siVIP) or with a Scrl sequence, and after 72 hr, the cells were harvested and used for RT‐qPCR. (a) Basal relative expression of VIP/VPACs is shown in BeWo cell line. Two highly positive controls (CT+) were used: the human epithelial colorectal adenocarcinoma cell line, CACO‐2 for VIP expression, and the human neuroblastoma cell line, SH‐SY5Y for VPAC receptors. Results are expressed as mean ± SEM of arbitrary units (A.U.) gene/housekeeping gene expression (n = 5). (b) The VIP expression in BeWo cell line transfected with VIP siRNA (siVIP) or scramble sequence (Scrl) were studied by qPCR. Results are expressed as mean ± SEM of fold change with respect to siVIP. P < 0.05 of seven independent experiments. (c) In the left panel, a double gradient heat map is shown: green for down‐regulated, black for no‐changes, and red for up‐regulated genes. In the right panel, the results are expressed as mean ± SEM of gene fold change (n = 5). In both, panels (b) and (c) the confidence interval was used to determine if the measurement in siVIP was statistically significant compared to Scrl. ECM, extracellular matrix; MMP, metalloproteinase; TIMP, tissue inhibitor of MMP; VIP, vasoactive intestinal peptide