Figure 6.

MYB4 mRNA and Protein Colocalize with SAD2 in Arabidopsis.

(A) Expression of MYB4, SAD2, C4H, CHS, CAD, and FAH1 in Arabidopsis. Total RNA was extracted from roots, leaves, stems, flowers, and siliques of 1-month-old wild-type plants. Real-time PCR analysis (40 cycles) was performed using gene-specific primers. Error bars indicate sd (n = 3).

(B) Coimmunoprecipitation of SAD2 and MYB4 proteins in vivo. The 2×35SP:6×myc-SAD2 plasmid was cotransformed into wild-type protoplasts with 35SP:3×flag-MYB4 or 35SP:3×flag-GL1. Total protein extracts were subjected to protein gel blot (WB) analysis with monoclonal anti-MYC (top panel) and anti-FLAG (middle panel) antibodies to detect the presence of myc-SAD2 and flag-MYB4/flag-GL1. Immunoprecipitation was performed using anti-c-MYC agarose conjugate. Bottom panel, protein gel blot analyzed with anti-FLAG antibody to detect coimmunoprecipitated flag-MYB4 and flag-GL1. Input, crude protein extract. Output, products pulled down by the anti-c-MYC agarose conjugate.