Figure 7. Dyn2 modulates granule secretion by NK clones.

A, NK cells were infected with recombinant vaccinia expressing WR or Dyn2. Cells were stimulated with plate bound anti-FcR or TPA (100 ng/ml) and ionomycin (10 µM). Supernatants were harvested after 4 hours at 37°C and granzyme A content was assessed using a BLT esterase assay. B, NK cells were electroporated with control or Dyn2 suppressing siRNA then stimulated using the indicated concentration of plate bound anti-FcR antibody. Additionally, low-dose PMA (10 ng/ml) and ionomyin (1 µM) were used to stimulate the Dyn2-suppressed cells. Granzyme A secretion was measured as described in A. Lysates were equalized for protein expression and blotted for Dyn2 to verify suppression of the protein. C, NK cells were treated with the indicated concentrations of dynasore and stimulated with plate-bound anti-FcR or TPA and Ionomycin. Granzyme A secretion was measured as described in A. D, NK cells were electroporated with control or Dyn2 suppressing siRNA and the percentage of NK cell – target conjugates expressing CD107a was determined using flow cytometry at the indicated time points.