bro1M4 and bro1M8 display WT steady-state membrane-associated ESCRT-III in vivo. Subcellular fractionation was performed in bro1Δ (GOY65) cells transformed with an empty vector or BRO1, bro1M4, or bro1M8 plasmids. vps4Δ (MBY3) cells were used as a control highlighting the distribution upon complete loss of Vps4 function. Representative immunoblots indicating fractionation of Snf7, Bro1, and Vps4 are shown. Pep12 and Pgk1 were used as membrane and soluble markers, respectively. Quantification represents six experiments; data are represented as mean ± SEM. Asterisks indicate a statistically significant difference compared with bro1Δ (P < 0.05).
Figure 8.

bro1M4 and bro1M8 display WT steady-state membrane-associated ESCRT-III in vivo. Subcellular fractionation was performed in bro1Δ (GOY65) cells transformed with an empty vector or BRO1, bro1M4, or bro1M8 plasmids. vps4Δ (MBY3) cells were used as a control highlighting the distribution upon complete loss of Vps4 function. Representative immunoblots indicating fractionation of Snf7, Bro1, and Vps4 are shown. Pep12 and Pgk1 were used as membrane and soluble markers, respectively. Quantification represents six experiments; data are represented as mean ± SEM. Asterisks indicate a statistically significant difference compared with bro1Δ (P < 0.05).

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