Figure 4.

The C-termini of both SND1 and PGAM5 mediate the binding of SND1 and PGAM5. (A) The diagrams show the structures of SND1 and PGAM5. (B) 293T cells were transfected with HA-PGAM5 plasmids alone or together with Flag-tagged plasmids expressing full-length SND1 (WT), its SN domains (N-terminus) or the TSN domain (C-terminus). Cell lysates were immunoprecipitated with anti-Flag antibody or IgG, followed by immunoblotting analysis with antibodies against HA and Flag. Actin served as loading control. (C) 293T cells were transfected with HA-SND1 plasmids alone or together with Flag-tagged plasmids expressing full-length PGAM5 (WT), the N-terminus, or the C-terminus. Cell lysates were immunoprecipitated with anti-Flag antibody or IgG, followed by immunoblotting analysis with antibodies against SND1 and Flag. Actin served as loading control. (D) GST pull-down of His-PGAM5 by GST-SND1 using protein purified from E. coli, followed by immunoblotting analysis with anti-PGAM5 and anti-GST antibodies.