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. Author manuscript; available in PMC: 2016 Dec 27.
Published in final edited form as: J Immunol. 2015 Oct 5;195(10):4760–4770. doi: 10.4049/jimmunol.1500979

FIGURE 5.

FIGURE 5

UNC-45A knockdown does not interfere with lytic granules polarization or with perforin expression levels. (A) YT NK cells transduced with either shRNA-scramble or shRNA targeting two different UNC-45A regions (1 and 2) were incubated with 721.221 target cells. After fixation, cells were stained with an anti-perforin mAb followed by Texas red goat anti-mouse IgG and analyzed by fluorescence microscopy. A representative experiment is shown. Arrows indicate NKISs. Scale bars, 5 µm. (B) Quantification of polarized granules per condition reported as the percentage of cells with polarized lytic granules out of the total number of conjugates. The averages of three independent experiments are shown. (C) Quantification of MFI in polarized regions in YT cell transduced with either scramble or shRNAs targeting UNC-45A regions (1 and 2). (D) Intracellular staining of perforin in YT cell transduced with either scramble of shRNAs targeting UNC-45A regions (1 and 2) or original, or treated with anti-mouse IgG as iso-type control and analyzed by FACS using a PE-conjugated anti-perforin Ab.