Fig. 3

Loss of Gimap5 results in impaired inactivation of GSK3β. a Association of GSK3β with vesicles in WT and Gimap5^sph/sph CD4⁺ T cells as exemplified by GSK3β intensity, vesicle number, and vesicle size of GSK3β spots 6 and 24 h after αCD3/αCD28 activation. b Bright detail similarity analysis (ImageStream) of activated CD4⁺T cells from WT and Gimap5^sph/sph mice show colocalization between Gimap5 and GSK3β. c Representative Z-stacks of WT and Gimap5^sph/sph CD4⁺ T cells stimulated for 24 h with αCD3/αCD28. d Immunoblot analysis of WT and Gimap5^sph/sph CD4⁺ T cells stimulated with αCD3/αCD28 depicting total and phosphorylated protein levels of GSK3β (P-Ser9 and P-Ser389), p38, and p53. e–i Localization of total and phospho-GSK3β (Ser389) in WT or Gimap5^sph/sph CD4⁺ T cells after 2 days stimulation with αCD3/αCD28 (n = 3). f Nuclear localization and h expression of total GSK3β. g Nuclear localization and i expression of p-GSK3β (Ser389). Hatched bars represent resting and solid bars represent CD3/CD28-activated CD4⁺ T cells. Graphs depict mean values ± SD. ImageStream data represent average values of >500 CD4⁺ T cells per sample. All experiments were performed at least three times. Statistical significance is determined by Student’s two-tailed test. BF bright field