Fig. 7

Hfl1 recruits Atg8 to vacuolar membrane to facilitate EVT. a The localization of Hfl1 on vacuolar membrane is regulated by its interaction with Atg8. Hfl1-GFP was expressed under the TEF1 promoter. In the absence of Atg8, or in the case of a mutant that loses the interaction (WIDY), Hfl1 was present in the vacuolar lumen in log-phase, with protein levels reduced in early stationary phase. Cells treated and image presented as in Fig. 1a. b Hfl1 promotes the association of Atg8 with vacuolar membrane. Both overexpression of Hfl1 and block of Atg8 delipidation enhanced the signal of GFP-Atg8 on the vacuolar membrane. Cells treated and image presented as in Fig. 1e. c Overexpression of Hfl1 does not bypass atg8Δ. Hfl1 was overexpressed using TEF1 promoter in WT and atg8Δ cells. Translocation of GFP-Pho8 was assessed as in Fig. 1a. d Forced lipidation of Atg8 bypass the need of Hfl1. Translocation of GFP-Pho8 was assessed as in Fig. 1a