Published customer image: . Goat anti Human Immunodeficiency Virus 1 gp120 antibody used for evaluation of envelope protein fragmants in HeLa cell lysates by western blotting. Image caption: . FIP1C depletion and HIV-1 Env incorporation in HeLa cells. (A) The efficiency of depletion of Rab11a-FIPs was measured by real time PCR (top). Infectivity of released particles following depletion of each FIP family member is shown below. (B) Depletion of FIP1C was confirmed at the protein level by immunoblotting with a FIP1C-specific antiserum. Env levels in both cell lysates and virions harvested from the FIP-depleted HeLa cells were assayed by immunoblotting. (C) Restoration of Env incorporation following shRNA-mediated depletion. Two constructs were used in this experiment: FIP1C cDNA is an shRNA sensitive GFP-tagged plasmid; while FIP1C* includes silent mutations in the shRNA target sequence rendering it shRNA-resistant. 2 ug and 4 ug of each FIP1C construct were used in the repletion assay as indicated at the top of the blot. (D) Infectivity of viral particles from the experiment shown in panel D as evaluated using TZM-bl indicator cells. Lanes are numbered and correspond to the blot above. Statistical comparisons utilized the unpaired t-test, * = p PLoS Pathog 9(4): e1003278.