TbLpn interacts with TbPRMT1. A) Far western analysis of TbPRMT1-TbLpn interaction. Purified His-TbLpn and His-RBP16 were separated on a 10% polyacrylamide gel, transferred to PVDF, and incubated with purified TbPRMT1 as described under Material and Methods. The blot was probed with anti-TbPRMT1 polyclonal antibodies (1:1,000), followed by goat anti-rabbit IgGs, and signals detected using chemiluminescence. B) In vivo interaction between TbLpn and TbPRMT1. TbLpn was immunoprecipitated from PF T. brucei cytosolic extracts using anti-TbLpn polyclonal antibodies as described under Material and Methods. As a negative control, the cytosolic extract was incubated in the absence of antibodies. Proteins present in the starting cytosolic fraction (C), as well as the bound (B) and unbound fractions (U) were separated on a 10% polyacrylamide gel and transferred to PVDF. The presence of TbLpn in the immune complexes was assessed by probing the membrane with anti-TbLpn polyclonal antibodies (1:1,000), followed by goat anti-rabbit IgGs. The presence of TbPRMT1 in the immune complexes was detected by probing the blot with anti-TbPRMT1 polyclonal antibodies (1:1,000), followed by goat anti-rabbit IgGs. Signals were detected using chemiluminescence.