Extended Data Fig. 2: Shigella flexneri efficiently inhibits caspase-3/-7 enzymatic activity without involving the cellular caspase inhibitor XIAP.
a, Caco-2, HEK293T, HUVEC or HDMEC cells were infected with ShigellaM90T, cytosolic lysates were extracted and lysates were treated with active caspase-8. Caspase-3 proteolytic processing and activity was analysed. Statistical significance to the corresponding non-infected value was determined by one-way ANOVA followed by Sidak’s post-analysis (n = 3 technical replicates of independent staining). b, CFU analysis of intracellular ShigellaM90T in Caco-2, HEK293T, HUVEC and HDMEC cells (n = 3 biologically independent samples). c, Western blot analysis of whole cell lysates derived from shScr and shXIAP HeLa cells. Asterisk mark unspecific band. d, CFU analysis of intracellular ShigellaM90T in shScr and shXIAP HeLa cells. Statistical significance to the corresponding shScr value was determined by one-way ANOVA followed by Sidak’s post-analysis (n = 3 biologically independent samples). e, Western blot analysis of whole cell lysates derived from HCT WT and XIAP-/- cells. Asterisk mark unspecific band. f, CFU analysis of intracellular ShigellaM90T in HCT WT and HCT XIAP-/- cells. Statistical significance to the corresponding WT value was determined by one-way ANOVA followed by Sidak’s post-analysis (n = 3 biologically independent samples). g, Live cell imaging of shScr or shXIAP HeLa cells infected with ShigellaM90T and treated with 100 ng ml−1 Trail. Scale bar, 100 µm. All experiments are representatives of at least two independent experiments and are presented as the mean ± s.d. **** p ≤ 0.0001; ** p ≤ 0.01. Weights in Western blots are in kDa.
