Fig 1. Integral membrane protein overexpression causes a growth defect in dfm1Δ cells in an ERAD independent manner.
(A) WT, dfm1Δ, and hrd1Δ cells containing either GALpr-HMG2-GFP or EV were compared for growth by dilution assay. Each strain was spotted 5-fold dilutions on glucose or galactose-containing plates to drive HMG2-GFP overexpression, and plates were incubated at 30°C. (B) Dilution assay as described in (A) except using WT, dfm1Δ, and doa10Δ cells containing either GALpr-STE6*-GFP or EV. (C) Dilution assay as described in (A) except using WT, dfm1Δ, and hrd1Δ cells containing either GALpr-PDR5*-HA or EV. (D) Dilution assay as described in (A) except using WT, dfm1Δ, and cdc48-2 cells. (E) Dilution assay as described in (B) except using WT, dfm1Δ, and cdc48-2 cells. (F) Dilution assay as described in (C) except using WT, dfm1Δ, and cdc48-2 cells. (G) Dilution assay as described in (A) except using WT or dfm1Δ cells expressing human CFTR, CFTRΔF508, or A1PiZ and plated only on glucose-containing plates. All dilution growth assays were performed in 3 biological and 2 technical replicates (N = 3). CFTR, cystic fibrosis transmembrane receptor; ERAD, endoplasmic reticulum-associated degradation; EV, empty vector.