Figure 9.

FoxP3 binds to nuclear ATF4 to hijack free ATF4 in cardiac remodeling (CR). (A-B) H9c2 cells and NRVMs grown on glass slides were treated with AngII (1 μM) with or without TP (10 μg/L) for 1 h. Then, the cells were fixed, blocked, and probed with antibodies against FoxP3 (green) and ATF4 (red). Confocal imaging and colocalization (yellow) were performed as shown in Figure 3I. (C-D) H9c2 cells grown in 10-cm dishes were treated as in Figure 3B for 1 h. Then, nuclear lysates were subjected to immunoprecipitation (IP) using an antibody against FoxP3, and the associated ATF4 was detected by immunoblotting (IB) or using an antibody against ATF4, and the associated FoxP3 was detected by IB. Protein ratios were calculated by normalization to histone H3 in the input (n = 3). (E-F) Experiments were performed as shown in C-D, and cytoplasmic lysates were used. Protein ratios were calculated by normalization to GAPDH in input (n = 3). ^*P < 0.05, ^**P < 0.01.