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. 2002 Sep 13;99(20):12859–12864. doi: 10.1073/pnas.192264999

Figure 4.

Figure 4

p38-mediated enhancement of IFN-γ-induced gene expression is STAT1-dependent and does not require phosphorylation of STAT1 at Ser-727. STAT1(−/−) MEFs reconstituted with STAT1-WT or -S727A mutant (A) or the parental STAT1(−/−) and the STAT1-WT reconstituted cells (B) were treated with anisomycin for 80 min (an) or with IFN-γ for 60 min (IFNg) or pretreated with anisomycin for 20 min followed by treatment with IFN-γ (60 min) (an/IFNg). Total RNA was isolated, and expression of IRF1 was determined in five (A) or two (B) independent experiments by using real-time RT-PCR carried out in triplicate or duplicate (a representative result is shown). The IRF1 value of each treatment was normalized to that of untreated cells (A Upper and B). Anisomycin-dependent enhancement is ≈3-fold in both STAT1-WT and -S727A cells, as shown by normalization of the values for the combined treatment with anisomycin (an) and IFN-γ (an/IFNg) to those of IFN-γ alone (IFNg) (A Lower). Note no significant induction of IRF1 in STAT1(−/−) cells by either treatment.