Plasma membrane depolarization activates the Rim101 pathway.
A, SEY6210 (WT) and YOK2098 (rim21Δ) cells harboring pFI1 (HA-RIM101) were grown at pH 5.5 and treated with CCCP or ethanol (mock) for 20 min. Immunoblotting was performed with anti-HA antibody or, to demonstrate uniform protein loading, with anti-Pgk1 antibody. B, total lysates were prepared from KCY662 (WT), YOK2098 (rim21Δ), KCY692 (lem3Δ), and KCY1113 (cho1Δ) cells grown in the presence of 1 mm ethanolamine. Immunoblotting was performed with anti-HA antibody or, to demonstrate uniform protein loading, with anti-Pgk1 antibody. FL, full-length; ΔC, Rim101ΔC.