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. Author manuscript; available in PMC: 2010 Oct 27.
Published in final edited form as: Immunity. 2009 Jul 17;31(1):60–71. doi: 10.1016/j.immuni.2009.05.010

Figure 3.

Figure 3

Mutational analysis of Vβ8.2, Vβ7 and Vβ2-containing iNKT TCRs. Staining of hybridomas expressing mutant versions of the Vα14i TCRα chain associated with the wild-type (A) Vβ8.2, (B) Vβ7 or (C) Vβ2 TCRβ chains. Staining of hybridomas expressing alanine substitutions of (D) Vβ8.2, (E) Vβ7 or (F) Vβ2 associated with the wild-type Vα14i TCRα chain. Dark blue, CDR1α; magenta, CDR2α; green, CDR3α, light blue, CDR1β and pink, CDR2β. WT, unsubstituted Vα14i TCRα chain paired with unsubstituted Vβ8.2, Vβ7 or Vβ2 TCRβ chains (wild-type controls). Vα3.2, Vα14i TCRα chain in which the CDR1α region is swapped for the Vα3.2 CDR1α region, and paired with the appropriate TCRβ chains (negative controls for TCRα substitutions). Vα13, Vα13-Jα18 TCRα chain paired with the appropriate TCRβ chains (negative controls for the TCRβ substitutions). ND, not done. The MFI of tetramer staining for each mutant was determined for a narrow TCR gate and normalized to wild-type MFI (set as 100%). Data represent the mean + s.e.m. of three independent experiments. Analysis of the Vβ8.2 mutants, with the exception of Y46A and E54A mutants, has been published previously in (Scott Browne et al., 2007) and is shown for comparison.