Fig. 4: SIRT2 is a potent lysine defatty-acylase that counteracts IcsB.
From: Golgi stress induces SIRT2 to counteract Shigella infection via defatty-acylation
A In-gel fluorescence detection of lysine fatty acylation of Flag-tagged IcsB substrate proteins treated with 5 μM of SIRT2, with or without 1 mM of NAD+ in vitro. B In-gel fluorescence detection of lysine fatty acylation of Flag-tagged IcsB substrates in HEK293T cells that were also transfected with Flag-tagged IcsB and SIRT2. Representative images from at least three independent experiments are shown. EV, empty vector. WT, SIRT2 WT. HY, SIRT2 HY. FL, fluorescence (indicative of lysine fatty acylation level on substrate proteins). Flag, anti-Flag immunoblot (indicative of input level of substrate proteins). C Quantification of the lysine fatty acylation levels in B. n = 4–6 biological replicates. D In-gel fluorescence detection of lysine fatty acylation of Flag-tagged Rac1 and Rac2 in HEK293T cells that were infected with equal number of WT or IpaJ deletion S. flexneri M90T cells. FL, fluorescence (lysine fatty acylation level). Flag, anti-Flag immunoblots. E Quantification of the lysine fatty acylation levels detected in D. n = 3 biological replicates. Data are represented as mean ± SEM. Statistical evaluation was done using one-way ANOVA. ***p < 0.001.
