Fig 1. LEF-11 exists in an oligomeric form.
(A) Oligomerization state of LEF-11, as determined by non-reducing SDS-PAGE analysis. BmN-SWU1 cells were transfected with pIZ-LEF-11cMYC plasmid. The cells were collected and lysed at 72 hours post transfection (h.p.t.). The samples were mixed with non-reducing protein loading buffer (without β-mercaptoethanol) without boiling, and α-cMYC antibody was added to detect oligomerization. +: with β-mercaptoethanol; -: without β-mercaptoethanol. (B) Co-immunoprecipitation (Co-IP) assays to verify LEF-11 homo-oligomerization. BmN-SWU1 cells were co-transfected with LEF11-FLAG and LEF11-cMYC plasmids. At 72 h.p.t., cells were collected and lysed for immunoprecipitation (IP) with α-FLAG antibody. Western blots were probed with α-FLAG antibody to detect the expression and immunoprecipitation of the input proteins and with α-cMYC antibody to detect co-immunoprecipitated LEF11-cMYC. Input, cell lysates; mouse IgG, IP with control mouse IgG.