Fig. 2.
HF activates the AAR pathway. (A) Histogram of microarray data from differentiating TH17 cells treated with HF or MAZ1310 for 3 or 6 hours. Red dots indicate transcripts increased greater than twofold by HF-treatment at both 3 and 6 hours. Text and arrows denote several defined AAR genes (27). (B) Quantitative real-time PCR was performed on cDNA from unstimulated T cells or those activated for 4 hours in the presence of MAZ1310 or HF. Asns, Gpt2, or eIF4Ebp1 mRNA expression was normalized to Hprt levels and are shown as mean values ± SD. (C) Unstimulated or TCR-activated T cells were treated with DMSO, 40 nM MAZ1310, or HF and were lysed for Western blotting after 4 hours. (D) Activated T cells were lysed at the indicated times after treatment with MAZ1310 or HF. ATF4 protein is indicated by an arrow. NS, nonspecific band. Microarray data were generated from three biological replicates. Other data represent two to three experiments.