Figure 5. Increased reactivity of TIPE2 knockout and knockdown macrophages to Toll-like receptor stimulation.
A. Bone marrow-derived macrophages from wild type and TIPE2-deficient mice (n=5) were treated with or without LPS (100ng/ml) for 8 hours. IL-6, IL-12p40 and TNF-α concentrations were determined by ELISA. B. TIPE2 mRNA levels in wild type and TIPE2 knockdown (KD) RAW 264.7 macrophages as determined by RT-PCR. GAPDH was used as a loading control whereas H2O was used as the background control. C–D. Wild type and TIPE2 knockdown RAW 264.7 cells were treated with LPS (100ng/ml) for the indicated times. IL-6 protein (C) and mRNA (D) levels were determined by ELISA and real-time PCR, respectively. E. Wild type and TIPE2 knockdown RAW 264.7 cells were treated with LPS (100ng/ml), peptidoglycans (PGN) (10μg/ml), Poly(I:C) (50μg/ml), or CpG oligodeoxynucleotides (CPG) (1μM) for 8 hours. IL-6 concentrations in the supernatants were determined by ELISA. Data shown are representative of three experiments. The p value shown is for all TLR treated WT cultures as compared to their respective TIPE2 KD groups. No significant differences were detected between the two groups in terms of the number and percentage of apoptotic cells as determined by annexin V staining.