Fig. 3.

Recruitment of CHMP4B by upstream ESCRT proteins. (A) Atto 488–labeled CHMP4B at 300 nM is recruited to Gag puncta by ΔPRD-ALIX at 4 μM. (B) Atto 488–labeled CHMP4B at 300 nM is recruited to Gag puncta by ESCRT-I (100 nM), ESCRT-II (200 nM), and CHMP6 (400 nM). (C) Same protein combination as in B except that ESCRT-II is omitted. (D) Quantification of CHMP4B recruitment to Gag puncta. Statistics of CHMP4B recrutiment was performed as for Fig. 2G, and the table shows added proteins analogously to Fig. 2G. Membrane is red, Gag is white, and fluorphore-labeled CHMP4B is green. Scale bar, 10 μm.