Nup format. Cell-free systems containing HeLa whole-cell lysates (WCL), cytosol, or cytosol plus isolated nuclei, were reacted with recombinant GST or GST-LE, in the presence or absence of recombinant His-Nup98 (A), or recombinant GST-Nup62 (B). After 45 min of incubation, the samples were fractionated, probed by Western analyses using α-Nup98 and α-His (A), or α-Nup62 (B). Pixel counts in the A, B, C, D boxes for each lane show the areas recorded and normalized to the control samples, as relative measures of observed phosphorylation.