FIGURE 5.

NAD-GAPDH during wheat seed development. Wheat seeds were collected at different DPA along development and the seed fresh weight (A) and starch content (B) determined immediately after sample collection. (C) NAD-GAPDH activity determined in total proteins extracted from each seed sample. (D) Immunoidentification of total NAD-GAPDH in whole protein extracts, phosphorylated NAD-GAPDH after phosphoprotein enrichment by IMAC-Fe³⁺ chromatography, SnRK1 and actin (used as a control) in whole protein extracts. (E) Immunoidentification of NAD-GAPDH in total wheat seed protein extracts at 27 DPA non-treated (lane 1) or treated (lane 2) with alkaline phosphatase before loading and after the IMAC-Fe³⁺ purification. In A–C, values are means of three independent assays determined by triplicate in two different biological samples. All immunoidentifications were performed in two different biological replicates and repeated independently twice in each replicate.