Supplementary Figure 6: Close contacts formed with an SLB.
From: Initiation of T cell signaling by CD45 segregation at 'close contacts'
(a) Montage from a time-lapse showing the spatio-temporal organization of CD45 (labeled with Alexa Fluor 488-tagged Gap 8.3 Fab, green) during contact of a Jurkat T cell expressing a transmembrane-anchored, non-signaling, T92A-mutated form of CD48 (CD48+) and a SLB containing rCD2 (Alexa Fluor 647-tagged, red). (b) Equivalent to (a) except that the fluorophores are swapped (CD45 is labeled with Alexa Fluor 647-tagged Gap 8.3 Fab, green, and rCD2 is tagged with Alexa Fluor 488, red). (c–e) Montages show distribution of a truncated form of CD45 (HA-CD45, c), TCR (d) and mLck (e) each labeled with HaloTag TMR, red) versus CD45 (green) during contact of a CD48+ Jurkat T cell with a SLB containing rCD2. The time intervals between frames shown are 5 s; data was collected at 37°C. (f) More examples of mLck versus CD45 distribution at T cell contacts with SLBs. Examples of rarer multi-focal contacts of CD48+ Jurkat T cells with rCD2-containing SLBs showing the organization of CD45 (labeled with Alexa Fluor 488-tagged Gap 8.3 Fab, green) and mLck labeled with HaloTag TMR (red).
