Figure 2.

2-AG effects on CHOP expression and modulation by cannabinoid signalling. Cells were treated with 2-AG (10 μM) for 24 h and expression levels of the pro-apoptotic ER-stress factor CHOP were assessed by qRT-PCR and Western blotting. For the assessment of CHOP dependency on cannabinoid signalling, cells were pre-incubated with the CB1 and CB2 antagonists AM281 and AM630, respectively. In BeWo cells, 2-AG treatment increases the transcript levels of CHOP encoded by the DDIT3 gene (A) and CHOP protein expression (B) through a CB2-dependent manner. Results show transcript levels normalised against GAPDH. A representative Western blotting and the densitometry analysis with relative ratios of CHOP/β-actin are shown, as β-actin was used as a loading control. (C) The increase in CHOP protein expression was also observed in cytotrophoblast cells isolated from term placenta and 2-AG effects were also reversed by CB2 receptor blockade with AM631, confirming the involvement of cannabinoid signalling through CB2 activation. A representative Western blotting and the densitometry analysis with relative ratios of CHOP/β-tubulin are shown, as β-tubulin was used as a loading control. Data are presented as the mean ± s.e.m. (*P < 0.05 vs control; ^#P < 0.05 vs 2-AG).