Fig. 2.

Coro1A is required for NK cell cytotoxic function but not activation. YTS cells were stably transfected with CORO1A-KD shRNA or EV. (A) YTS-EV or YTS-CORO1A-KD cells were lysed and fractionated by high-speed centrifugation; cytoplasmic (C) and insoluble pellet (P) fractions were then immunoblotted for actin and Histone H1 as a loading control. (B) YTS-EV (solid line) or YTS-CORO1A-KD (dashed line) cells were incubated with susceptible 721.221 target cells for 1 h (Left) or 4 h (Right), and cell lysis was assayed by standard ⁵¹Cr-release assay. The mean ± SD of four independent experiments is shown. *P < 0.05 by Student's two-tailed unpaired t test. (C and D) Conjugates were formed as described in Materials and Methods in the presence of anti-CD107a to assess NK cell degranulation. YTS-EV (black) cells were normalized to 1. (E) YTS-EV (WT) or YTS-CORO1A-KD (KD) cells were activated for 10 min on anti-CD18 and anti-CD28 or mouse IgG as a negative control. Cells were immunoblotted for phosphorylated ERK or Myosin IIA as a loading control. Shown is one representative blot from three independent experiments.