Fig. 9.

ACOT11 and ACOT1 knockdown selectively perturbs mitochondrial and cytosolic S-deacylase activities. After transfection with either control or ACOT11 or ACOT1 shRNA vectors, HEK293T treated with 1 µM Hoechst 33342, 100 nM MitoTracker Deep Red for 30 min, washed, loaded with 1 µM DPP-2 (a, e) or 500 nM mitoDPP-2 (c, g) for 10 min, and then analyzed by fluorescence microscopy. b Quantification of experiment shown in (a). ACOT11 perturbation does not affect DPP-2 (cytosolic) signal. d Quantification of experiments shown in (c). ACOT11 knockdown causes an increase in mitoDPP-2 (mitochondrial) signal. f Quantification of experiment shown in (e). ACOT1 knockdown causes an increase in DPP-2 (cytosolic) signal. h Quantification of experiments shown in (g). ACOT1 perturbation does not affect mitoDPP-2 (mitochondrial) signal. For all imaging, 20 µm scale bar shown. For all plots, statistical analyses performed with a two-tailed Student′s t-test with unequal variance, *P value < 0.05; **P value < 0.007, n = 5 for (a, c, e and g), error bars are ± s.e.m