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. 2013 Feb 21;121(17):3386–3395. doi: 10.1182/blood-2012-11-468868

Figure 4.

Figure 4

Silencing SAP or Fyn in DN3A4-1.2 iNKT cells results in reduced cytotoxicity. (A-B) DN3A4-1.2 (1.2) cells were stably transduced with lentiviruses encoding control (Ct), SAP- (S1, S2; [A]), or Fyn-specific (F1, F2; [B]) shRNA. After 5 days, cell lysates prepared from untransduced (WT) or shRNA-transduced cells were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotted with anti-SAP or Fyn antibodies to assess knockdown of SAP or Fyn expression, respectively. (C-F) The in vitro cytolytic activity of untransduced 1.2 NKT cells or 1.2 cells transduced with control, SAP-specific (C-D), or Fyn-specific (E-F) shRNA was tested against PBS44-loaded EL4 cells. Data are from 1 of 4 experiments. (G-J) Mean percent reduction ± SEM in cytolysis of EL4 cells by SAP or Fyn-silenced 1.2 iNKT cells compared with WT cells (S1 or S2; [G-H] and F1 or F2; [I-J]). Data are averaged from 4 experiments. Statistical significance in percent-specific lysis of PBS44-loaded EL4 cells by SAP- or Fyn-silenced 1.2 iNKT cells compared with WT cells was determined by two-way ANOVA test. **P < .001.