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note
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2 level of vehicle treated-control)] × 100
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25
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note
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2 dishes for 24 h. Then, compound 1, or 7 was treated with 1 μg/mL of LPS for 6 h. After washing with PBS, total RNA was isolated from the cells. Briefly, cells were lysed by Tri-reagent and separated into three phases by chloroform. The aqueous phase was transferred to a fresh tube, and added the same amount of isopropanol to precipitate RNA pellet. RNA pellet was washed with 75% ethanol and dissolved in nuclease free water. The concentration of RNA was determined by absorbance at 260 nm. RNA (1 μg) was performed reverse transcription (RT) using avian myeloblastosis virus (AMV) reverse transcriptase. The cDNA products of RT were amplified by PCR using Taq DNA polymerase. The sense and antisense primers for COX-2 were 5′-GGAGAGACTATCA-AGATAGTGATC-3′ and 5′- ATGGTCAGTAGAC-TTTTACAGCTA-3′, respectively. The sense and antisense primers for β-actin were 5′-TGTGATGG-TGGGAATGGGTCAG-3′ and 5′-TTTGATGTCA-CGCACGATTTCC-3′, respectively. The PCR products were run on a 2% agarose gel and visualized by SYBR Gold staining
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