Fig 1: Disparate kinetics of gene inactivation and protein loss for DOCK2, CD90, CD49d/integrin a4 and CCR7 in naïve CD8+ T cells. (A) Decay of normalized MFI values of target proteins among targetlo cells. (B) Relative MFI values of target proteins calculated by normalizing the values presented in (A) to the MFI value on day 0. Curves defined as y = [100 – (Plateau)] × exp(–K × x) were fitted to the data, with R2 = 0.979, 0.997, 0.999 and 0.999 for DOCK2, CD90, CCR7 and CD49d, respectively. Plateau values and half-life were computed during curve fitting. (C) Kinetics of relative knockout efficacy calculated by normalizing %DOCK2-GFPlo and %CD90lo in each experiment to the corresponding values on day 10. Data were fitted with the equation y = [Plateau] × (1 – exp(–K×x)], with R2 = 0.984, 0.970, 0.999 and 0.926 for DOCK2, CD90, CCR7 and CD49d, respectively. Plateau value and half-time to reach the plateau was estimated from the data during curve fitting. Results of ordinary one-way ANOVA with Dunnett’s multiple comparison of differences in half-life shown in (B, C) are summarized in Table 2. Graphs show pooled data from 3–6 independent experiments. Values of %CD90lo from experiments 5 and 6 (as indicated in Figure 7) were excluded in C because of their continuous increase until day 10.