Real-Time PCR
Real-Time PCR procedure was carried out in thermal cycler Roche
LightCycler® 480 Real-Time PCR System (Roche Diagnostics Ltd, Basel,
Switzerland) with the use of Power-Up SYBR Green Master Mix (Applied
Biosystems, California, USA). The target genes in our study wereCNR1 , CNR1a, and CNR2 encoding CB1, CB1a, and CB2,
respectively. The reference genes in the procedure were ACTB (β-actin)
and GAPDH (glyceraldehyde 3-phosphate dehydrogenase). In a single
reaction, we used 1 µl of cDNA. Gene-specific primers were used at a
10µM concentration (Table 1).