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. 2003 Mar;23(5):1750–1763. doi: 10.1128/MCB.23.5.1750-1763.2003

FIG. 5.

FIG. 5.

Coprecipitation of Far3 with Far7 to Far11. Strains containing myc-tagged Far7 to Far11 were subjected to IP with a polyclonal rabbit HA antibody. Proteins contained in the IP were separated on duplicate SDS-PAGE gels and analyzed by Western blotting using monoclonal mouse antibodies to detect Far3-HA and myc-tagged Far7 to Far11. Pre-IP lysates were also analyzed by Western blotting to detect myc-tagged Far proteins or the loading control, Dpm1. Each myc-tagged Far protein was tested in isogenic strains with the following alterations: far3 mutant plus empty vector (YEp352), far3 mutant plus high-copy-number Far3-HA (pSL2784), and wild-type plus plasmid-borne HA tag (pSL2771). Far10-myc and Far11-myc were also tested in a far3 far9 background that contained the high-copy-number Far3-HA plasmid (pSL2784). Lane letters indicate strain details and experimental conditions, as follows: A, far3 〈YEp352〉 + IP α-HA (polyclonal rabbit anti-HA antibody); B, FAR3 〈pSL2771〉 + IP α-HA; C, far3 〈pSL2784〉 no IP α-HA; D, far3 〈pSL2784〉 + IP α-HA; E, far3 far9 〈pSL2784〉 + IP α-HA; X, 1:2 dilution of the concentrations in panel D. Strains used in each lane were as follows: Far7-myc (A, SY4065; B, SY4078; C and D, SY4066); Far8-myc (A, SY4067; B, SY4079; C and D, SY4068); Far9-myc (A, SY4069; B, SY4080; C and D, SY4070); Far10-myc (A, SY4071; B, SY4081; C and D, SY4072; E, SY4077); Far11-myc (A, SY4073; C, D, and X, SY4074; E, SY4084).