Fig. 7

Evolution of pHi in different yeast cell inocula (CEN.PK 113-5D expressing pHluorin, TMB3800) upon exposure to a mixture of lignocellulosic inhibitors. Representative histograms from the following inocula: a LP-cells, b cells pre-adapted in medium supplemented with the inhibitors at pH 5, and c ESP-cells. The vertical line displays cells with high and low ratio (F405/F488) and was defined as the number (=0.400) separating LP-cells (a, inoculum) into fractions of 1% lowest and 99% highest percentiles. d Mean ratio (F405/F488) and e frequency of cells with high ratio (>0.400) for the three different inocula (LP-cells, grey bars; pre-adapted cells, blue bars; ESP-cells, green bars) measured over time. Error bars represent standard error of biological duplicates (n = 2). Ratio (F405/F488) is the ratio of pHluorin fluorescence emission collected at 542/50 and 533/30 nm originating from excitation at 405 and 488 nm, respectively. Ratiometric flow cytometry measurement was performed with a Moflo cell sorter and data analysis was performed with Flowjo