Analysis of CB₁R and CB₂R expression in resting and activated primary human T-lymphocytes.

<p>Freshly isolated T-lymphocytes were either left untreated or treated with 5 µg/ml anti-CD3 and 2 µg/ml anti-CD28 for the indicated periods of time (24–72 hours). <b>Panels A–D</b>: Cells were lysed and the homogenates were assayed for CB₁R and CB₂R protein by Western blot analysis. CB₁R and CB₂R protein levels were quantified by densitometric analysis, and were normalized to beta-actin. The results were expressed as total amount of receptors over 24 hours and 72 hours of stimulation (<b>A</b>, <b>B</b> and <b>C</b>) (average value of all samples) as well as CB₂R to CB₁R ratio (<b>Panel D</b>). Panel <b>E</b>: Cells were washed in PBS, and then mRNA was isolated and analysed by qRT-PCR using specific primers for CB₁R and CB₂R. Data are representative of 4 independent experiments.