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. Author manuscript; available in PMC: 2012 Apr 21.

Published in final edited form as: Cell Host Microbe. 2011 Apr 21;9(4):286–298. doi: 10.1016/j.chom.2011.03.012

hGas6 enhances infection of the EEV preparation of vaccinia virus. (A) HMVEC were incubated with PBS (+) or PBS (+) containing hGas6 (100 ng/ml). The IMV form was used for infection at MOIs of 0.1, 0.5 and 2.5. The EEV preparation was used for infection at undetermined MOIs. (B) HMVEC were incubated for 1 hour with PBS (+) or PBS (+) containing either normal goat IgG or goat anti-human Axl antibody (25 µg/ml) as a blocking reagent then incubated with hGas6 (100 ng/ml) in the presence of one blocking reagent for 4 hours. The EEV preparation (undiluted) was incubated for 1 hour with the same volume of PBS (+) or PBS (+) containing ANX5 (25 µg/ml) and applied to HMVEC pretreated with hGas6 and blocking reagent. (C) 293T and Axl-293T cells were incubated with AIM-V or AIM-V containing hGas6 (100 ng/ml), followed by infection with the EEV preparation (100-fold diluted supernatant). (A–C) EEV preparations were pretreated with anti-A27L antibody to neutralize possible contamination of IMV (Figure S4) (Ichihashi, 1996).

hGas6 enhances infection of the EEV preparation of vaccinia virus. (A) HMVEC were incubated with PBS (+) or PBS (+) containing hGas6 (100 ng/ml). The IMV form was used for infection at MOIs of 0.1, 0.5 and 2.5. The EEV preparation was used for infection at undetermined MOIs. (B) HMVEC were incubated for 1 hour with PBS (+) or PBS (+) containing either normal goat IgG or goat anti-human Axl antibody (25 µg/ml) as a blocking reagent then incubated with hGas6 (100 ng/ml) in the presence of one blocking reagent for 4 hours. The EEV preparation (undiluted) was incubated for 1 hour with the same volume of PBS (+) or PBS (+) containing ANX5 (25 µg/ml) and applied to HMVEC pretreated with hGas6 and blocking reagent. (C) 293T and Axl-293T cells were incubated with AIM-V or AIM-V containing hGas6 (100 ng/ml), followed by infection with the EEV preparation (100-fold diluted supernatant). (A–C) EEV preparations were pretreated with anti-A27L antibody to neutralize possible contamination of IMV (Figure S4) (Ichihashi, 1996).