Fig. 3.

Tissue response to S. Typhi infection reflects differential apical cytokine release.

(A) Multiplexed ELISA analysis of pro-inflammatory and regulatory cytokines in control biopsies, STM or STY treated (black, red, blue, respectively).

(B) As cytokine release is observed but no transcriptional upregulation is detected (per RNA-seq data set), intracellular signal transduction was assessed by western blot analysis to determine the phosphorylation state of NF-kB and MAPK from samples mounted in the microsnapwell system after exposure to media, STM or STY at time points of 60 min and 120 min.

(C) Densitometry analysis of signaling blots.

(D) Pathway analysis demonstrates a central role for MAPK in regulated cellular processes and gene expression important in development of immune responses. Note: All genes shown here are downregulated and their expression patterns predict inhibition of cell recruitment.

(E) NF-kB is a central regulator of gene expression; its activation is critical to expression of genes downregulated in our RNAseq data set.