IFN Regulatory Factors 4 and 8 Expression in the NOD Mouse
Figure 4
Reduced expression of IRF4 and IRF8 in tolerogenic BMDCs derived from NOD mice. Total RNA was extracted from BM-DCs generated with GM-CSF or a combination of GM-CSF and IL-4, purified and gene expression profiles of IRF4 (a) and IRF8 (d) were analyzed by RT-PCR. Each sample was analyzed in triplicates and normalized to HPRT expression (ΔCt). Each NOD sample was normalized to BALB/c sample (ΔΔCt = ΔCt sample − ΔCt BALB/c), and the expression levels were calculated as follows: 2−ΔΔCt. Data are shown as the average ±SD of 3-4 independent experiments (* and, **). (b) and (e) total proteins were extracted from BM-DCs generated with GM-CSF or a combination of GM-CSF and IL-4 and analyzed by Western blotting for IRF4 (b) or IRF8 (f) expression. The relative intensities of bands were assessed using the NIH Image software and normalized to reference actin bands to establish a ratio of (c) IRF4/actin and (f) IRF8/actin. The expression levels observed in BALB/c mice were arbitrarily set as a unitary value. A representative of 2 independent experiments is shown. Error bars correspond to S.D.
