Phosphorylation of STATc is reduced in the absence of Pyk3 and Phg2.

<p>Ax2 wild-type, pyk3⁻, phg2⁻, and pyk3⁻/phg2⁻cells were either treated (+) for 15 minutes with 200 mM sorbitol (A), 20 mM 8-Br-cGMP (B), 30 µM BHQ (C) or left untreated (−). Total cell lysates were prepared, proteins were separated by 10% SDS-PAGE and transferred to nitrocellulose. Total and tyrosine phosphorylated STATc were detected with the CP22 and 7H3 antibodies, respectively. (D–F): Quantification of phosphorylated STATc in either treated or untreated cells. Band intensities were determined densitometrically and normalised using the values for total STATc. The amount of phosphorylated STATc of treated Ax2 cells was set to 1 and relative values were calculated for untreated Ax2 cells as well as treated and untreated mutants. The error bars depict standard deviations of three (D, E) and six (F) independent experiments. (G–I): Staple diagrams depicting the relative amount of phosphorylated STATc from treated (black part) versus untreated (white part) cells. The total amount of phosphorylated STATc was set to 1 for each comparison and relative values were calculated.