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. Author manuscript; available in PMC: 2013 Oct 15.
Published in final edited form as: J Immunol. 2012 Sep 12;189(8):4088–4103. doi: 10.4049/jimmunol.1200777

Figure 9. Requirement for de novo mRNA and protein synthesis in mA3 mRNA induction.

Figure 9

(A and B) Raw cells were stimulated with 1000 units/ml of endotoxin-free recombinant IFNα, 1g/ml LPS, or vehicle. Stimulant-containing medium was removed and replaced with fresh medium with or without 1ug/ml Actinomycin D (ActD). Cells were collected every hour for 15 hours as indicated on the figures. Total RNA was extracted and used for cDNA synthesis and cDNA used for examination of level ofmA3 mRNA by qPCR. (C and D) Raw cells were pre-treated with 10μg/ml CHX for 30 minutes followed by stimulation with 1000 units/ml of endotoxin-free recombinant IFNα, 1μg/ml LPS, or vehicle. Cells were collected at the time points indicated on the figure for total RNA was extraction and cDNA synthesis. Level of mA3 transcript was examined by qPCR. In addition, STAT1 and STAT2 mRNA levels were examined. Data is normalized to GAPDH and presented as fold change relative to vehicle treated cells. Error bars are standard error. Experiments were repeated at least three different times with similar results.