FIG 2.

Requirements of cellular factors for LUJV VLP production. (A, B) Tsg101 and/or ALIX/AIP1 were depleted by siRNA treatment, and their effects on LUJV (A) or LASV (B) Z-mediated VLP production were examined. Endogenous Tsg101 and ALIX/AIP1 were detected with specific antibodies. The LUJV and LASV Z proteins were detected with anti-FLAG antibody. Normalized Tsg101 and ALIX protein levels are shown at the bottom. (C, D) LUJV and LASV VLPs produced by pC-LUJV-Z-FLAG or pC-LASV-Z-FLAG with overexpression of DN (Vps4AEQ and Vps4BKQ) and WT Vps4A/B forms were examined by WB. Overexpressed DN and WT forms of Vps4A/B, LUJV Z, and LASV Z were detected with anti-FLAG antibody. Normalized VLP production (VLPs per cell) is shown on the right. Actin (used as a loading control) was also detected in the experiments whose results are shown in panels C and D. The data shown are averages and standard deviations of three independent experiments.