Figure 1

From: Cannabinoid 2 receptor induction by IL-12 and its potential as a therapeutic target for the treatment of anaplastic thyroid carcinoma

Figure 1

(A) Real-time reverse transcription (RT)-PCR analysis of cannabinoid 2 receptor (CB2) mRNA in un-transfected ARO cells, ARO cells transfected with vector (ARO/vector) or interleukin (IL)-12 plasmid (ARO/IL-12) and ARO cells incubated with 10 ng ml−1 IL-12 for 16 h (ARO+IL-12). The data are expressed as fold increase of CB2 mRNA in ARO/IL-12 cells vs ARO cells. (B) Northern analysis of CB2 gene expression in ARO and ARO/IL-12 cells. The differential expression of CB2 gene between the two cell lines was verified by northern blot hybridization of 1.2 kb CB2 cDNA probe to 20 μg of total RNA extracted from both cell lines (upper panel). The actual RNA loading was monitored by ethidium bromide staining of RNA loaded for northern analysis (lower panel). (C) Confocal microscopy analysis of CB2 protein in ARO and ARO/IL-12 cells: (a) ARO cells stained with goat anti-rabbit secondary antibody alone (background control); (b) ARO/IL-12 cells stained with polyclonal rabbit anti-CB2 antibody and goat anti-rabbit secondary antibody and (c) ARO cells stained with polyclonal rabbit anti-CB2 antibody and goat anti-rabbit secondary antibody. (D) Expression of IL-12 receptor B1 and B2 subunits in ARO and ARO/IL-12 cells. Both subunits were amplified by RT-PCR using primers specific for B1 and B2 subunits. The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a control. M: 100 bp ladder; lanes 1 and 2: ARO and ARO/IL-12 RNA amplified for 168-bp IL-12 B1 fragment; lanes 3 and 4: ARO and ARO/IL-12 RNA amplified for 275-bp IL-12 B2 fragment; lanes 5 and 6: ARO and ARO/IL-12 RNA amplified for 300 bp GAPDH.