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[Preprint]. 2024 Mar 25:2023.11.13.566830. Originally published 2023 Nov 13. [Version 2] doi: 10.1101/2023.11.13.566830

Figure 5. Individual amino acid substitutions that have occurred in gorilla CD4 protect against SIVcpz entry.

Figure 5.

(A,B,D,E) HIV-1ΔEnv-GFP viruses were pseudotyped with Envs from diverse SIVcpz isolates (MB897, blue; EK505, orange; MT145, green; TAN1.910, purple). Cf2Th cells stably expressing human CCR5 and wild-type (wt) or mutated human or gorilla CD4 (X-axis) were infected with these pseudoviruses and then the percent cells infected (GFP-positive cells) were enumerated by flow cytometry 48 hours post infection. Data represent the mean +/− SEM from two independent experiments, each with two technical replicates. Stars above data sets signify that both independent experiments showed significant statistical differences (p < 0.05) when compared to wild-type by one-way ANOVA. (C) Lysates of Cf2Th cells stably expressing the indicated CD4 receptors in “A” and “B” were treated with PNGase F (to remove N-specific glycans) or left untreated and then probed for CD4 expression by western blotting. The number of N-specific glycosylation sites within the D1-domain of CD4 was determined computationally (Gupta and Brunak, 2002) and is shown under the blot. β-Actin served as a loading control. Chimpanzee CD4, with two D1-domain glycans, has been described previously (Warren et al., 2019b).