Fig. 4.
Applicaton of NHE-3, NaPi-IIa, NHE-1, and MAP17 to the MYTH 2.0 system. TF-Cub-Baits (pTLB-1) derived from full-length NHE-3 or its C-terminal truncation for THM (ΔTHM) (A), NaPi-IIa or its C-terminal truncation for TRL (ΔTRL) (B), NHE-1 (C), and MAP17 (D) were co-expressed with the following NubG-HA-Preys: PDZK1, NHERF-1, NHERF-2, CAL, NaPi-IIa, NHE-3, Ost1-NubG (negative control), and Ost1-NubI (positive control). Reconstitution of split Ub was determined by yeast growth on minimal SD−Trp/−Leu (SD−WL) medium. The interactions of baits with preys were analyzed by growth on selective SD−Trp/−Leu/−Ade/−His (SD−WLAH) medium supplemented with 3-AT (left and middle panels) as described in Fig. 2B. Quantification of MYTH reporter activity stemmed from ONPG liquid tests on permeabilized cells and was depicted against Miller units to normalize cell densities (right panels). The error bars show the standard deviations determined from triplicates. PM, plasma membrane.