FIGURE 7.

Effects of IFN-γ and TSA on pSTAT-1/IRF-1 protein expression and IFN-γ-inducible gene transcription in mouse TBCs. A, RNA was isolated from SM9 and M-11 cells exposed to 500 U/ml IFN-γ, 50 nM TSA, or the combination, for 24 h and subjected to SYBR Green-based quantitative RT-PCR using primers specific for IRF-1, GBP, LMP-2, and GAPDH, as described for Fig. 2. The data are the average of four independent experiments and are represented as the ratio of the relative mRNA expression of each gene (i.e., IRF-1) vs GAPDH. Unpaired Student’s t test was used to compare the levels of mRNA expression in SM9 and M-11 cells treated with the combination of IFN-γ/TSA vs IFN-γ alone. B, WCE were prepared from NIH-3T3, SM9, and M-11 cells that were treated as described in A, and subjected to WB analyses, using Abs specific for STAT-1, phosphotyrosine (701)-STAT-1, and IRF-1. The blots were stripped and reprobed with Abs for HSC70 as a control for loading and the integrity of the protein extracts. Representative data from three independent preparations of WCE are shown.