Figure 1.

Workflow of iTRAQ, 2-DE, Western blot, and MRM of the urinary proteome. For analysis of the urinary proteome, 3 iTRAQ experiments were performed, 2-DE, Western blot, and MRM were conducted to confirm and validate the iTRAQ results. iTRAQ experiments 1, 2, and 3 were performed, labeled (a) and (b), (c) and (d), and (e), respectively, wherein 3 biological replicates (labeled (a), (d), and (e), resp.), technical replicate 1 (labeled (a) and (c)), and technical replicate 2 (labeled (b) and (d)) were performed in microalbuminuric versus normoalbuminuric urine. 2-DE, Western blot, and MRM analysis of the urinary proteome were conducted using labeled (f), (g), and (h), respectively.