FIG 10.
Endocytosis and trapping of artificial Env in ERC. FAP-TMCT is an artificial envelope bearing the FAPalpha2 module at the N terminus fused to the TM and CT of HIV-1 Env. (A) FAP-TMCT labeled following a 10-min pulse with MG-11p on ice, showing a z section at approximately the midpoint of the cell. (B) Same cells as those described for panel A, showing labeling at the PM (coverslip level). Nuclei are superimposed for reference. (C) PM labeling on ice in HeLa cells expressing GFP-FIP1C560–649. (D) HeLa cell expressing GFP-FIP1C560–649 and pulse labeled with MG-11p for 5 min, shown immediately at the end of the pulse. (E) FAP-TMCT colocalization with GFP-FIP1C560–649 after a 5-min pulse followed by a 20-min chase period. (F) FAP-TMCT colocalization with GFP-FIP1C560–649 following a 5-min pulse and 60-min chase period. (G to I) Control membrane protein produced from pMFAP-alpha2 vector using methods described for panels D and E. (J to L) FAP-TMCT144 time course with time postlabeling indicated. Scale bars, 11 μm. (M) Pearson's correlation was derived from image stacks representing 10 fields of cells from each time point. Error bars indicate standard deviations.