Extract total RNA from at least 2 x 10^6 cells using TRIZOL reagent and an RNeasy RNA extraction kit. 2. Reverse-transcribe 60 ng of total RNA with a High-capacity cDNA Reverse Transcription kit. 3. Prepare a reaction mix with 12.5 μl 2X Sybr Green PCR Master Mix, 2.5 μl 1 μM primer mix, 2 μl cDNA (5 ng/μl), and 8 μl water. 4. Perform Real-Time PCR on 10 ng of cDNA using a Power Sybr Green PCR Master Mix with the following protocol: 50°C (2 min), 95°C (10 min), 95°C (15 sec), 60°C (1 min), for 40 cycles. 5. Conduct a dissociation assay to evaluate primer specificity. 6. Analyze data with built-in SDS Analysis Software and adjust conditions if issues are detected before repeating the assay.