Effects of MPA treatment on the interaction of TIF-IA and Ebp1 and the regulation of PCNA expression in T cells. (A-B) Effects of MPA on the interaction of TIF-IA and Ebp1 and PCNA levels in cultured primary T cells. Cells were treated with MPA for 24 hours. (A) Immunoprecipitation with anti-Ebp1 antibody (far left), immunoblotting with the antibodies indicated (near left), and PCNA expression and MTS assay (near right and far right). (B) Effects of the T340N mutation on the interaction of TIF-IA and Ebp1 and PCNA expression. Jurkat T cells were transfected with Myc–TIF-IA or Myc–T340N–TIF-IA and GFP Ebp1. Immunoprecipitation with anti-GFP antibody (far left), expression of p53 and PCNA (near left), PCNA mRNA expression (near right), and MTS and colony forming assay (far right). (C) Interaction of TIF-IA and Ebp1 and expression of PCNA in primary T cells from controls or MPA-treated individuals. Primary T cells were isolated from PBMCs of healthy donors (n = 6) or MPA-treated patients (n = 5). Lysate was incubated with anti-Ebp1 antibody and the precipitate blotted for TIF-IA. (D) PCNA mRNA expression.