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. 2003 Aug;23(15):5388–5400. doi: 10.1128/MCB.23.15.5388-5400.2003

FIG. 1.

FIG. 1.

Cdc34 self-associates in vivo. (A) Cross-linking. Cell lysates from YPH499 yeast cells expressing Flag-Cdc34 were treated with or without the chemical cross-linker DSS. Cell lysates were subsequently analyzed by immunoblotting (IB) with an anti-Flag antibody. The position of Flag-Cdc34 is indicated, as is the position of a unique cross-linked product containing Flag-Cdc34 (arrow). (B and C) Coimmunoprecipitation. Total cell extracts of YPH499 cells expressing Flag-Cdc34, Myc-Cdc34, or both Flag-Cdc34 and Myc-Cdc34 were prepared. (B) Myc-Cdc34 was immunoprecipitated (IP) with an anti-Myc antibody, followed by immunoblotting with an anti-Myc antibody (right panel) to detect the amount of Myc-Cdc34 that had immunoprecipitated and with an anti-Flag antibody (left panel) to detect the amount of Flag-Cdc34 that had coimmunoprecipitated. (C) The reciprocal coimmunoprecipitation experiment to that for panel B was performed. Flag-Cdc34 was immunoprecipitated with an anti-Flag antibody, followed by immunoblotting with an anti-Flag antibody (right panel) and with an anti-Myc antibody (left panel). The position of Cdc34 is indicated, as is the position of a proteolytic product of Cdc34 (*). Protein expression levels within the lysates used for immunoprecipitations are shown at the bottoms of panels B and C.