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. 2012 Sep 17;198(6):1025–1037. doi: 10.1083/jcb.201202137

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© 2012 Andrés-Delgado et al.

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Figure 4. — FH2 domain expression allows Glu-MT formation and MTOC polarization in DIA1, FMNL1, or INF2 KD cells. (A and B) Jurkat cells were transfected with DNA constructs coexpressing GFP and shDIA1 or shFMNL1. Cells were immunoblotted for Dia1, FMNL1, or GAPDH (A). The histogram represents the percentage of DIA1 or FMNL1 content (B). (C and D) DIA1-, FMNL-1 (C), or INF2-KD cells were left untransfected or were transfected with constructs expressing the indicated FH2 domains (D). After conjugation to SEE-loaded APCs, cells were stained for Glu- and Tyr-MTs. The arrowheads indicate the position of the MTOC of the T cells. The histogram represents the percentage of T cells with Glu-MTs or polarized MTOC (D). Bars, 5 µm. At least 40 T cells were analyzed in D. Data in B and D are summarized as means ± SEM from three independent experiments (error bars; *, P < 0.05; **, P < 0.01; ***, P < 0.001).