CKIδ regulates TCR-mediated centrosome polarization to the IS. (A) Conjugates of Jurkat and SEE-pulsed Raji cells in the presence of the indicated drugs. Centrosomes are stained with anti–γ-tubulin antibody. Raji cells are blue in merge. The box plot shows quantification of centrosome polarization to the IS based on scoring criteria in Fig. S1 A (n = 4 experiments; 200 conjugates/experiment). (B) Conjugates of Jurkat and SEE-pulsed Raji cells in the presence of the indicated drugs. Cells are stained with anti–PY174-Vav and anti–γ-tubulin antibodies. Raji cells are blue in merge. (C) Cytoplasmic cell extracts of Jurkat cells containing stably integrated EV (control), vector encoding CSNK1D shRNA (clones shdelta1 and shdelta2), or CSNK1E shRNA (clone sheps1) are immunoblotted with antibodies against CKIδ or CKIε. Actin serves as a loading control. WB, Western blotting. (D) Conjugates of control (EV), CKIε- or CKIδ-depleted Jurkat cell clones, and SEE-pulsed Raji cells. Centrosomes are stained with anti–γ-tubulin antibody. Raji cells are blue in merge. The box plot shows quantification of centrosome polarization to the IS (n = 4 experiments; 200 conjugates/experiment). In the box plots, whiskers are set at minimum and maximum, and horizontal lines mark the median, whereas boxes indicate the interquartile range (25–75%). Bars:10 µm; (en face) 5 µm.