Fig. 2.

Spontaneous reexpression of the TbbHpHbR gene restores susceptibility to TLF-1 in susceptible T. b. brucei 427–060^R. (A) Third round of FACS for T. b. brucei 427–060^R cells that bound Alexa Fluor–488–conjugated TLF-1. Successive rounds of sorting enriched for a subpopulation of cells that bound TLF-1, T. b. brucei 427–060^S. T. b. brucei 427–221^S minus TLF-1 is blue; T. b. brucei 427–221^S plus TLF-1 is black; T. b. brucei 427–060^R plus TLF-1 is green; and T. b. brucei 427–060^R (third FACS sort) plus TLF-1 is orange. SI Materials and Methods provides details of FACS. (B) Analysis of Alexa Fluor-488–conjugated TLF-1 uptake by FAC. T. b. brucei 427–221^S plus TLF-1 is black; T. b. brucei 427–060^R plus TLF-1 is green; clonal line from the TLF-1–positive subpopulation in the third round of FACS T. b. brucei 427–060^S plus TLF-1 is red. (C) In vitro TLF-1 lysis assay: percentage of cells lysed following incubation with TLF for 2 h at 37 °C. T. b. brucei 427–221^S is marked as a black triangle; T. b. brucei 427–060^R a green square; T. b. brucei 427–060^S a red circle. (D) Northern blot analysis of total RNA with probes specific for VSG 060, TbbHpHbR (Hp/Hb receptor), and β-tubulin mRNAs. Lane 1, T. b. brucei 427–060^R; lane 2, T. b. brucei 427–060^S.