Fig. 9
From: A non-conserved amino acid variant regulates differential signalling between human and mouse CD28
Vav SH2 domain is not required for Nck recruitment to CD28 and actin rearrangement and polarisation. a Human CD28WT cells were transfected with pm-Cherry Vav1 WT or Vav R696A mutant (magenta) together with GFP-Nck WT (green) and then stimulated for 15 min in the absence (Ctr) or presence of Dap3/B7 cells (B7). After fixing and permeabilisation F-actin was stained with 633-conjugated phalloidin (blue) and analysed by confocal microscopy. The scale bar represents 10 μm. The frequency of conjugates (total 15) showing significant (RRI > 2.5) Vav1 and Nck recruitments (b, c) or F-actin polarisation (c) were calculated and data represent the mean ± SD of three independent experiments. Mean values: Vav WT, Nck = 5.1 ± 2.5, Vav = 4.4 ± 1.7, F-actin = 7.1 ± 2.9; Vav R696A, Nck = 4.3 ± 1.2, Vav = 3.3 ± 1.4, F-actin = 6.7 ± 3.1. NS not significant
