preTCR complex is stabilized at cell surface at lower temperatures. (A) Plots show the individual unnormalized datasets used to construct Fig. 4 B. (B) Flow cytometry of surface-stained HEK cells stably expressing either the αβTCR or preTCR complexes, when incubated at either 37°C (solid line) or 25°C (dashed line). Filled histogram denotes staining of untransduced HEK cells. (C) Flow cytometry plots showing surface staining of αβTCR as a function of expression when transfected HEK cells are rapidly transitioned from 25°C to 37°C. Indicated time denotes the period since temperature shift, with the final plot showing αβTCR-expressing HEK cells maintained at 37°C. Dataset is representative of three biological replicates. (D) Equivalent dataset as in B for the preTCR complex. (E) Relative mean surface staining of αβTCR and preTCR expressed in HEK cells as a function of time after temperature shift. Datapoints have been normalized to the initial value when cells are maintained at 37°C and are shown accumulated from three biological replicates.