Figure 4.

Reciprocal Purification of Augmin Subunits and Identification of Two Plant-Specific Subunits.

(A) Immunoaffinity purification results using transgenic plants expressing AUG3-c-myc, AUG4-c-myc, or AUG5-c-myc. Purified proteins together with those from a wild-type negative control are revealed in a silver-stained gel. Asterisks indicate the bait for each purification. The positions of AUG1, 3, 4, and 5 are indicated at the right side of the gel image. M, molecular weight markers.

(B) Immunoblotting analysis using anti-c-myc, anti-AUG1, anti-AUG3, anti-AUG4, and anti-AUG5 antibodies. Proteins purified from AUG3-c-myc (A3), AUG4-c-myc (A4), and AUG5-c-myc (A5) include all subunits of the augmin complex. The baits (asterisks) and endogenous augmin subunits (arrows) are revealed by immunoblotting. WT, wild type.

(C) Mass spectrometry results obtained through three independent immunopurifications using AUG3-c-myc, AUG4-c-myc, and AUG5-c-myc as baits. Besides AUG1-6, two additional subunits (AUG7 and AUG8) were identified. Gene ID represents gene identification number by TAIR; # peptides, number of unique peptides; # spectra, number of total spectra; coverage, sequence coverage. The bait of each experiment is highlighted in gray.