*2.6. Western Blot*

Western blots of placental lysates were done following standard procedures, as previously described [25]. As primary antibodies, we employed the following: rabbit polyclonal antibodies

anti E-cadherin (Cat. 3195, Cell Signaling Technology, Inc., Danvers, MA; 1:3000); anti claudin-1 (Cat. 51–9000, Invitrogen, Camarillo, CA; dilution 1:1000); anti claudin-2 (Cat. 51–6100, Invitrogen, Camarillo, CA; dilution 1:1000); anti claudin-3 (Cat. Ab52231, Abcam, Cambridge, MA; dilution 1:500); anti claudin-4 (Cat. 36–4800, Invitrogen, Camarillo, CA; dilution 1:4000); anti claudin-5 (Cat. ab15106, Abcam, San Diego, CA; dilution 1:2000); anti claudin-7 (Cat. 34–9100, Invitrogen, Camarillo, CA; dilution 1:500); anti occludin (Cat. 71–1500, Invitrogen, Carlsbad, CA; dilution 1:1000); anti ZO-1 (Cat. 61–7300, Invitrogen, Carlsbad, CA; dilution 1:1000); and anti ZO-2 (Cat. 71–1400, Invitrogen, Carlsbad, CA; dilution 1:500). We also employed mouse monoclonal antibodies anti JAM-C (Cat. sc-515893, Santa Cruz Biotechnology, Santa Cruz, CA; dilution 1:500) and anti-JAM-B (Cat. sc-293496, Santa Cruz Biotechnology, Santa Cruz, CA; dilution 1:500). As secondary antibodies, we employed goa<sup>t</sup> antibodies against rabbit IgG coupled to peroxidase (Cat. 6111620, Invitrogen, Camarillo, CA; dilution 1:20,000), or against mouse IgG coupled to peroxidase (Cat. 626520, Invitrogen, Camarillo, CA; dilution 1:10,000).
