Figure 5.

Quantitative proteomic analysis of N. benthamiana leaves transiently expressing 3×Flag-SlREM1. A, The experimental workflow for quantitative proteome analysis. Total proteins were extracted from N. benthamiana leaves transiently expressing 3×Flag-SlREM1 (SlREM1) and 3×Flag (control check [CK]) and subjected to iTRAQ analysis. The four different colors used oin the right representation different labeling reagents (115, 116, 117, and 118) used in the iTRAQ experiment. B, Total of 272 proteins showing differential expression were identified in N. benthamiana leaves transiently expressing 3×Flag-SlREM1 (SlREM1) compared to the CK. Proteins showing significant changes in abundance were hierarchically clustered based on the abundance ratio as a log₂ scale. Each row in the color heat map represents a single protein. The yellow and red colors represent upregulated and downregulated proteins, respectively, in the N. benthamiana leaves transiently expressing 3×Flag-SlREM1 relative to the control. Values represent the average of three biological replicates. The differentially regulated proteins are listed in Supplemental Table S2. C, The number of proteins showing higher or lower abundance in N. benthamiana leaves transiently expressing 3×Flag-SlREM1 relative to the control at 1 and 3 dpi. D, The overlapped proteins identified at 1 and 3 dpi in N. benthamiana leaves transiently expressing 3×Flag-SlREM1 relative to the control. E, Identification of proteins involved in cell death, ROS generation and scavenging, defense response, and lignin biosynthesis in quantitative proteome. Gray box indicates that the protein is not identified at 1 d. Abundance ratios were calculated using CK as denominator and plotted in a heat map. Values represent the average of three biological replicates.