Figure 2
Serine 181 phosphorylation is dynamically regulated by Src activity. (a) Western blot analysis of levels of ZBP1 phosphorylated at Ser181 (P-ZBP1[S181]) and total ZBP1 in protein lysates obtained from Neuro2a cells transfected with pEGFP-ZBP1, pEGFP-ZBP1Y396F, and pEGFP-ZBP1GxxG. GAPDH is shown as an additional loading control. Cropped blots are presented (for full length blots see Supplementary Information). (b) Results of quantitative Western blot analysis. The data are presented as a ratio of the signal intensity of P-ZBP1(S181) to the signal intensity of ZBP1 ± SEM. *p < 0.05, ***p < 0.001 (One sample t-test). Number of independent experiments (N = 3). (c) Western blot analysis of levels of ZBP1 phosphorylated at Ser181 (P-ZBP1[S181]), total ZBP1, and Src in protein lysates obtained from Neuro2a cells transfected with pBIO-ZBP1 and plasmids that encoded SrcWT, SrcCA, or SrcDN. GAPDH is shown as an additional loading control. Cropped blots are presented (for full length blots see Supplementary Information). (d) Results of quantitative Western blot analysis. The data are presented as a ratio of the signal intensity of P-ZBP1(S181) to the signal intensity of ZBP1 ± SEM. *p < 0.05, ns - non significant (One sample t-test). Number of independent experiments (N = 4).
