Taqman primers for mouse immune response

Ischemic hemispheres and were harvested from mice 96 hours after MCAO, flash frozen in liquid nitrogen and stored at −80°C until processed. Spleens were processed as described above and 1×10⁶ cells were flash frozen in liquid nitrogen and stored at −80°C until processed. Total RNA was isolated using the RNeasy Mini Kit according to the manufacturer’s instructions. (Qiagen, Valencia, CA). cDNA was synthesized using the SuperScript II Reverse Transcriptase cDNA synthesis kit (Life Technologies, Grand Island, NY). Quantitative real-time PCR was performed using the StepOnePlus Real-Time PCR System with TaqMan primers for mouse immune response (Life Technologies, Grand Island, NY). Primers used are as follows: IL-6 (Mm00446190_m1), TNFα (Mm00443258_m1), IL-1β (Mm00434228_m1), iNOS (Mm00440502_m1), MMP-9 (Mm00442991_m1), CD206 (Mm01329362_m1), ICAM-1 (Mm00516023_m1), VCAM-1 (Mm01320970_m1), p-Selectin (Mm01295931_m1). GAPDH housekeeping gene was used as an endogenous control. Results were analyzed using ExpressionSuite Software (Thermo Fisher Scientific, Waltham, MA).

Ischemic hemispheres and were harvested from mice 96 hours after MCAO, frozen in liquid nitrogen and stored at −80°C until processed. Spleens were processed as described above and 1×10⁶ cells were frozen in liquid nitrogen and stored at −80°C until processed. Total RNA was isolated using the RNeasy Mini Kit according to the manufacturer's instructions. (Qiagen, Valencia, CA). cDNA was synthesized using the SuperScript II Reverse Transcriptase cDNA synthesis kit (Life Technologies, Grand Island, NY). Quantitative realtime PCR was performed using the StepOnePlus Real-Time PCR System with TaqMan primers for mouse immune response (Life Technologies, Grand Island, NY). Primers used are as follows: IL-6 (Mm00446190_m1), TNFα (Mm00443258_m1), IL-1β (Mm00434228_m1), iNOS (Mm00440502_m1), MMP-9 (Mm00442991_m1), CD206 (Mm01329362_m1), ICAM-1 (Mm00516023_m1), VCAM-1 (Mm01320970_m1), p-Selectin (Mm01295931_m1). GAPDH housekeeping gene was used as an endogenous control. Results were analyzed using ExpressionSuite Software (Life Technologies, Grand Island, NY).