Expression and molecular cloning of the mouse IL12RB1 gene. (A) IL-15 and IFN-γ, but not LPS, induce IL12RB1 gene expression in a mouse macrophage cell line. RAW264.7 cells were left untreated (unstim) or treated for 4 hours with 1 μg/mL LPS, 10 ng/mL IL-15, or 10 ng/mL IFN-γ. Total RNA (20 μg each) was prepared for the Northern blot analysis using a 32 P-labeled IL12RB1 cDNA probe. Gene expression of IL12RB1 and a picture of the ethidium bromide (EtBr)–stained gel are shown. (B) Transcription start sites of the IL12RB1 gene were mapped by primer extension analysis. The transcribed product (P) was run along with sequencing ladder from the same primer on a 6% polyacrylamide/urea gel. Asterisks indicate the transcribed products; 3 transcriptional start sites are shown, and the proximal major site was designed as +1. (C) Schematic presentation of the mouse IL12RB1 gene. Black boxes represent exons. Positions of 3 transcriptional start sites are indicated.
