Blood Journal
Leading the way in experimental and clinical research in hematology

Kurowska M, Goudin N, Nehme NT, et al. Terminal transport of lytic granules to the immune synapse is mediated by the kinesin-1/Slp3/Rab27a complex. Blood. 2012;119(17):3879-3889.

On page 3887 in the 26 April 2012 issue, the right portion of Figure 6C did not publish as intended. Some of the dots, indicating the number of vesicles/cell for both KIF5B and KIF5 DN, are missing. The corrected Figure 6 is shown.

Figure 6

Kinesin-1 regulates the terminal transport of the Rab27a/Slp3 granules at the IS. (A) Confocal microscopy of CTLs transfected with GFP-KIF5B, DsRed-Slp3, and Flag Rab27a and conjugated with L1210 target cells (top panel, insets represent an enlarged image). Confocal microscopy of CTLs transfected with GFP-KIF5 DN, DsRed-Slp3, and Flag Rab27a and conjugated with L1210 target cells (bottom panel, insets represent an enlarged image). All images of conjugates were representative of > 90 cells observed over at least 3 independent experiments. (B) CTLs were transfected with CFP-Slp3, CFP-KIF5B, and Flag-Rab27a or CFP-Slp3, CFP-KIF5 DN, and Flag-Rab27a. Transfected cells were incubated for 3 hours alone or with a coated anti-CD3 antibody in the presence of CD107a and CD107b antibodies. The cells were then stained with anti-CD3–PE and anti-CD8–APC. Lymphocytes were gated on CD3+CD8+. Profile shows CFP versus CD107-FITC gating on CD3+CD8+ CTLs. For each condition, we perform a histogram overlay analysis of Lamp secretion by CFP+ cells. Each condition is representative of 2 independent experiments performed in duplicate. (C) Representative TIRF images of mCherry-KIF5B/GFP-Rab27a/GFP-Slp3 and mCherry-KIF5DN/GFP-Rab27a/GFP-Slp3 transfectant CTLs. The “pseudocolor” scales were used to indicate the intensity staining in TIRF. A vesicle was defined by a minimum of a square of 3 pixels per 3 pixels (1 pixel = 0.16 μm) of strong intensity signal (orange to purple). Scale bars represent 5 μm. The number of vesicles per cell acquired in TIRF mode is shown on the graph. Each dot represents an individual cell; and the line, the median value. For the mCherry-KIF5B/GFP-Rab27a/GFP-Slp3 condition, 20 cells have been analyzed from cumulating 3 independent experiments. For the mCherry-KIF5DN/GFP-Rab27a/GFP-Slp3 setting, 16 cells have been analyzed from cumulating 3 independent experiments. Statistical analysis was performed using a Mann-Whitney test. ***P < .0001.