Figure 4. Effect of LAMP-1 truncation on TMEM175 inhibition.
(A) Constructs of the two LAMP-1 deletion mutants. Brackets mark the deleted regions.
(B) Sample I-V curves and Cs+ current density (at 100 mV) of HEK293 cells expressing TMEM175 with LAMP-1 or its truncation mutants. The Cs+ currents were recorded in whole-cell configuration at pH 7.4. Data are represented as mean ± SEM (n=10, ** p <0.01).
(C) Sample I-V curves and proton current density (at −100 mV) of TMEM175 in complex with LAMP-1 or its truncation mutants. The proton currents were recorded with pH 5.5 in the bath and pH 7.4 in the pipette. NMDG+ was used as monovalent in both bath and pipette solutions. Data are represented as mean ± SEM (n=10, ** p <0.01).
(D) K+ flux assay using proteoliposomes containing TMEM175, TMEM175/LAMP-1 before or after de-glycosylation (DG), or TMEM175/LAMP-1-TM.