Inhibition of Triggering Receptor Expressed on Myeloid Cells 1 Ameliorates Inflammation and Macrophage and Neutrophil Activation in Alcoholic Liver Disease in Mice

Tornai, David; Furi, Istvan; Shen, Zu T.; Sigalov, Alexander B.; Coban, Sahin; Szabo, Gyongyi

Total RNA was extracted using the Qiagen RNeasy kit (Qiagen) according to the manufacturer's instructions with on-column deoxyribonuclease treatment. RNA was quantified using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific), and complementary DNA synthesis was performed using the iScript Reverse Transcription Supermix (Bio-Rad Laboratories) and 1 µg total RNA. Real-time quantitative polymerase chain reaction (PCR) was performed using Bio-Rad iTaq Universal SYBR Green Supermix and a CFX96 real-time detection system (Bio-Rad Laboratories). Relative gene expression was calculated by the comparative ∆∆Ct method. The expression level of target genes was normalized to the housekeeping gene 18S ribosomal RNA in each sample, and the fold change in the target gene expression among experimental groups was expressed as a ratio. Primers were synthesized by IDT, Inc.; the sequences are listed in Table 1 .