Bro1^ΔBOD does not support efficient MVB cargo sorting of Mup1, Mup1-Ub, and Cos5. This figure complements Fig. 2. (A) Representative immunoblots showing processing of Mup1-GFP or Mup1-GFP-Ub, including liberated GFP in WT (SEY6210), vps4Δ (MBY3), bro1Δ (GOY65), bro1^ΔBOD (bro1Δ::TEF1p- bro1^ΔBOD ; CTY2), bro1^ΔBOD snf7Δ (CTY12), bro1^ΔBOD vps4Δ (CTY5), or bro1^ΔBOD doa4Δ (CTY13) cells. Pgk1 serves as loading control. (B) Representative immunoblots processing of Cos5-GFP, including liberated GFP in WT (SEY6210), vps4Δ (MBY3), bro1Δ (GOY65), bro1^ΔBOD (bro1Δ::TEF1p- bro1^ΔBOD ; CTY2), bro1^ΔBOD snf7Δ (CTY12), or bro1^ΔBOD vps4Δ (CTY5) cells. Pgk1 serves as loading control. Data were quantified from three independent experiments performed on three separate days from two transformations. Data are represented as mean ± SD, Asterisks indicate a statistically significant difference (P < 0.05), and number signs indicate statistically significant difference compared with WT (P < 0.0001). (C) Lysates generated from bro1Δ (GOY65), WT (SEY6210), and TEF1p-bro1^ΔBOD (CTY2) cells expressing GFP-Dap2 were analyzed by immunoblotting using antibodies against GFP and Pgk1 (loading control).