Figure 3. Cellular proteins progressively downregulated by HIV infection.
(A) Enlargement of cluster #1 from hierarchical cluster analysis (Figure 2A). The heatmap shows log2 (fold change in protein abundance compared with uninfected cells). Enzymes associated with deoxynucleotide metabolism (blue) and B56 family regulatory subunits of serine/threonine protein phosphatase PP2A (red) are highlighted, along with known Vif target APOBEC3C (boxed) and other proteins of interest (bold). (B) Temporal profiles of enzymes associated with deoxynucleotide metabolism (blue) and B56 family regulatory subunits of serine/threonine protein phosphatase PP2A (red). Relative abundance is expressed as a fraction of maximum TMT reporter ion intensity, and the temporal profile of APOBEC3C is shown for comparison.
Figure 3—figure supplement 1. Immunoblot validation of novel HIV targets.
Depletion of proteins in cluster #1 by HIV-1 infection. CEM-T4s were infected with NL4-3-dE-EGFP HIV at an MOI of 10 and analysed by immunoblot (IB) 48 hr post-infection. Depletion of positive control (CD4 and APOBEC3G) and novel (RRM2 and PPP2R5A/D) HIV targets was confirmed. ImageJ (Schneider et al., 2012) was used to determine band intensities, which were normalised to calreticulin intensity and compared with TMT-based proteomic quantitation at 48 hr. p24 (capsid) and Vif are included as additional controls.