Skip to main content
. 2006 Mar 15;34(5):e41. doi: 10.1093/nar/gkl053

Figure 5.

Figure 5

(A) Quantitative cell-to-cell fusion assay. NIH-3T3 cells transfected with pCMV-α and the corresponding pMSCV-GaLV construct were mixed with an equal number of HEK-293T target cells expressing pCMV-ω and incubated at 37°C for either 5 h (dark bars) or 22 h (bright bars). Subsequently, β-galactosidase activity in cell lysates was determined using a luminometric assay. (B) R peptide cleavage of selected variants and controls. Cell lysates of transfected HEK-293T cells were subjected to immunoprecipitation with an anti-HA antibody followed by western blot analysis using anti-TM antibodies. Untransfected cells served as a negative control (lane 9). As positive control for R peptide cleavage, purified MLV particles having the wild-type GaLV Env incorporated were loaded in lane 1. The positions of the unprocessed (TM) and processed (TM-ΔR) form of the TM protein are indicated. The light chains of the anti-HA antibody used for immunoprecipitation are labeled by an asterisk. Arrows point to the positions of the selected TM protein variants.