Cell - 8 September 2016

(D) Neutralization activity detected in the serum of GLHL121 and MutHGLL121 mice immunized according to the regimen described in (C). Purified Igs from the
serum of immunized GLHL121 and MutHGLL121 mice were tested for neutralization activity in a TZM-bl assay against a panel of 14 Tier 2/1B HIV-1 isolates. R1166
naturally lacking the N332 glycan was used as a control for epitope specificity.
(E) Graphs show the results of ELISAs performed with the serum (1/100 dilution) of naive or immunized GLHL121 and MutHGLL121 mice using the sequential
protocol indicated on top. ELISAs against the immunogens at different steps of the immunization protocol are shown.
(F) Neutralization activity detected in the serum of GLHL121 and MutHGLL121 mice immunized according to the regimen indicated in (E). Purified Igs from the
serum of immunized GLHL121 and MutHGLL121 mice were tested for neutralization activity in a TZM-bl assay against a panel of 4 Tier 2/1B HIV-1 isolates. Murine
Leukemia Virus (MuLV) was included as a negative control. IC 50 < 0.096 in red; 0.096- 0.5 in orange; 0.5- 4 in yellow; > 4 green; not detectable (ND) in gray; not
tested indicated by a dash. When indicated in the graphs, the specificity of the serologic response to the PGT121 epitope was evaluated by ELISA against the
10MUT and 10MUT KO proteins.