Dithiothreitol (dtt)

All reagents were from Sigma, except where indicated, with the following stock compositions. Yeast nitrogen base (Difco); Choline (1 M stock in DDW); Rapamycin (GoldBio; primary stock solution: 2.5 mg/ml in 100% ethanol, and secondary stock of 1 mg/ml in 90% ethanol+10% Tween‐20 (v:v), final 200 ng/ml). Copper sulfate (used at 500 μM from a 0.5 M stock in DDW). Potassium Permanganate (Dissolved in DDW), S‐1 (or Dimethylethanolamine)—Ted‐Pella−18315. BEEM embedding capsules size 00, EMS, 70010‐B. Proteinase K (stock 2 mg/ml, final 100 mg/ml (diluted in PS200)); Triton X‐100 (stock 40% (w/v), final 0.4% (w/v)); DTT (Bio Basic Inc); PMSF (200 mM in EtOH); Z1000 Zymolyase 20T (US biological); Sodium‐Sulfate (BDH); Mouse anti‐GFP (MBL); mouse anti‐Pgk1 (Abcam); Rabbit anti‐Ape1 (polyclonal antibodies prepared by using two synthetic peptides that were conjugated to keyhole limpet hemocyanin (KLH) and then injected into rabbits to produce an anti‐Ape1 antiserum that recognizes both precursor and mature forms); Anti‐cpy1 polyclonal antibody (obtained from Prof. Howard Riezman); pRS316‐GFP‐Atg8 (expressed under the promoter of Atg8, kind gift from Prof. Yoshinori Ohsumi); pDP105 (expressing TagBFP‐Ape1 under the native Ape1 promoter and Ape1 under the pCUP1 promoter, kind gift from Prof. Claudine Kraft).

Strains, plasmids and oligonucleotides used in this study are listed in Tables S1–S4. All C. albicans strains were maintained on YEPD medium (1% yeast extract, 2% bacto peptone, 2% glucose and 2% agar for solidification). Prototropic transformants were selected on a synthetic medium (2% dextrose, 1.7% Difco yeast nitrogen base with ammonium sulfate and auxotrophic supplements). For nourseothricin‐resistant mutants, 200 μg ml⁻¹ of nourseothricin (Werner Bioagents, Jena, Germany) was added to the YEPD. To obtain nourseothricin‐sensitive transformants, strains were grown in YPM (1% yeast extract, 2% peptone and 2% maltose) for 8 hr and plated on YPM plates containing 25 μg ml⁻¹ nourseothricin for 48 hr. Except for sodium dodecyl sulfate (SDS; Biobasic, Inc.) and DTT (SRL), all the supplements and chemicals; β‐mercaptoethanol, tunicamycin, fluconazole, ketoconazole, voriconazole, amphotericin B, calcofluor white and congo red were purchased from Sigma‐Aldrich. Caspofungin was obtained from Merck & Co. Inc., NJ.