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. 2011 Mar 28;108(15):6085–6090. doi: 10.1073/pnas.1015328108

Fig. 3.

Fig. 3.

Arginine methylation of BAD inhibits its phosphorylation by Akt. (A) BAD peptides (amino acids 87–103) that were unmodified control (Ctrl) or methylated at Arg-94 and/or Arg-96 (meR94, meR96, and meR94/meR96) were incubated with or without recombinant Akt1 in the presence of [32P]ATP. The extent of BAD phosphorylation was measured using a scintillation counter. Values are means ± SD of three independent experiments. (B) HEK293T cells were transfected with FLAG-BAD together with or without wild-type or G98R mutant of HA-PRMT1. Whole-cell lysates were immunoprecipitated with anti-FLAG antibody, and then the immunopurified FLAG-BAD was treated with bacterial alkaline phosphatase, followed by incubation with recombinant Akt1 in the presence or absence of ATP. Reaction products were analyzed by immunoblotting as indicated. (C) HEK293T cells were transfected with FLAG-BAD with or without the wild-type or G98R mutant of HA-PRMT1. Whole-cell lysates were analyzed by immunoblotting as indicated. (D and E) Whole-cell lysates from HEK293T cells transfected with control (Ctrl) or PRMT1-specific siRNA were analyzed by immunoblotting as indicated.