Figure 3.

Cannabinoid receptor CB₂ in embryonal carcinoma. (A) Expression of CB₁ and CB₂ receptors in Nt2d1 cell line. Mouse brain and spleen were used as positive controls for the expression of CB₁ and CB₂, respectively. (B) Analysis of the activation of ERK pathway following the stimulation with 1 µM JWH-133 at the indicated time points. In (A) and (B), tubulin was used as loading control. (C) Cell cycle profile of Nt2d1 cells in untreated and JWH-133 treated cells (1 µM), at the indicated time point. The sub-G1 pick is indicative of nuclear fragmentation. (D) Chronic exposure of Nt2d1 cells at 1 µM JWH-133 causes an arrest at the G1/S phase of the cell cycle. (E) Expression of CB₂ receptor following chronic exposure of Nt2d1 cells at 1 µM JWH-133 for the indicated time frames. The expression of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control. (F) Densitometric analysis of CB₂ expression treatment of Nt2d1 cells with 1 µM JWH-133 for 72hs. CB₂ expression was normalized against the loading control (GAPDH). Data are mean value ± s.d. of three independent experiments. Statistical analysis was performed using a unpaired two-tail Student’s t-test (p < 0.05; Barchi and Grimaldi, unpublished data).