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. Author manuscript; available in PMC: 2016 Mar 3.
Published in final edited form as: Biochemistry. 2015 Feb 19;54(8):1681–1693. doi: 10.1021/bi501171r

Figure 2. Effects of alanine substitution on HXB2 Env mediated cell-cell fusion and viral entry.

Figure 2

For cell-cell fusion (black bars), each mutant plasmid was transfected into 293T cells along with plasmids expressing the tat and rev proteins. The transfected cells were then co-incubated with the receptor cell line TZM-bl for 24 hours. Fusion activity was measured by luciferase assay. For viral entry (white bars), each mutant was transfected into 293T cells along with the HIV backbone plasmid and virus was harvested after 48 hours. The amount of virus was determined by a p24 antigen capture ELISA assay, and virus stocks with an equivalent amount of p24 protein (100 ng) were added to each well of TZM-bl cells. Viral entry levels were determined by luciferase assay at 48 hours post infection. The WT level and the 50% of WT fusion are indicated with a dotted line.