Supplemental Figures s2

SUPPLEMENTAL FIGURES

Figure S1 – Replication Kinetics of Vif-proficient and Vif-deficient HIV-1IIIB on

Panels of Human APOBEC3-Expressing SupT11 T Cell Lines

(A) The replication kinetics of Vif-proficient (gray diamonds) and Vif-deficient

(black squares) HIV-1IIIB were monitored over three weeks of a spreading infection on panels of SupT11 clones stably expressing each APOBEC3 protein by periodic infection of the reporter line CEM-GFP. The results for four clones from each panel

are shown here. Vif-proficient HIV-1IIIB replicates with similar kinetics on permissive

(APOBEC3-low) cell lines, such as CEM-SS and SupT11, and nonpermissive

(APOBEC3-high) cell lines, such as CEM and H9. Vif-deficient HIV-1IIIB only replicates on permissive cell lines. Stable expression of A3A, A3B, or A3C in the permissive

SupT11 line results in no inhibition of Vif-deficient HIV-1IIIB replication relative to

Vif-proficient HIV-1IIIB. On the other hand, Vif-deficient HIV-1IIIB replication is significantly delayed or undetectable on cell lines expressing A3D, A3F, A3G, or A3H.

In cell lines expressing moderate to high levels of A3H, Vif-proficient HIV-1IIIB

replication is also undetectable or delayed, reflecting the inability of HIV-1IIIB Vif to completely neutralize C-terminally tagged A3H as previously reported (4, 5) and as in Figure 2C.

(B) Immunoblots of the SupT11 clones, whose infectivity plots are shown above, demonstrate stable expression of each HA-tagged APOBEC3. The order of the infectivity plots corresponds to the loading order of the samples. Figure S2 – Replication Kinetics of Vif-proficient and Vif-deficient HIV-1IIIB on

Panels of Rhesus APOBEC3-Expressing SupT11 T Cell Lines

(A) The replication kinetics of Vif-proficient (gray diamonds) and Vif-deficient

(black squares) HIV-1IIIB were monitored over three weeks of a spreading infection on panels of SupT11 clones stably expressing each rhesus APOBEC3 protein by periodic infection of the reporter line CEM-GFP. The results for three clones from each panel are shown here. Stable expression of rhesus A3A, A3B, or A3C in the

permissive SupT11 line results in no inhibition of Vif-deficient HIV-1IIIB replication

relative to Vif-proficient HIV-1IIIB similar to the vector control lines. On the other

hand, Vif-deficient HIV-1IIIB replication is undetectable on cell lines expressing

rhesus A3D, A3F, A3G, or A3H. Vif-proficient HIV-1IIIB replication is similarly suppressed on rhesus A3F and A3H lines and is delayed on rhesus A3G lines,

reflecting the inability of HIV-1IIIB Vif to completely neutralize these proteins.

(B) Immunoblots of the SupT11 clones, whose infectivity plots are shown above, demonstrate stable expression of each HA-tagged rhesus APOBEC3. The order of the infectivity plots corresponds to the loading order of the samples.