ZFIN Image: Jim et al., 2022, Figure 1

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Figure Caption

Figure 1

Dual RNA sequencing of host-pathogen transcriptomes in early pneumococcal meningitis

(A) Zebrafish meningitis infection model. Pneumolysin-deficient S. pneumoniae D39V (PLY−) mutant bacteria or complemented S. pneumoniae D39V (PLY+) bacteria (∼2,000 CFUs) were injected into the hindbrain ventricle of 2 dpf zebrafish larvae.

(B) Bacterial load in zebrafish larvae infected with S. pneumoniae PLY+ or S. pneumoniae PLY− at 8 hpi. The data represent the mean ± SD of two biological replicates with 9–10 larvae per group; each dot represents a single larva; ns, non-significant; determined by unpaired t test.

(C) Total RNA was isolated from pooled infected zebrafish larvae (n = 100 per biological replicate) for preparation of cDNA libraries and sequencing at 8 hpi. Scale bar: 100 μm.

(D) Quality control (QC) was performed on raw reads, low-quality reads were trimmed, and remaining reads were aligned to a synthetic chimeric genome. Aligned reads were counted and classified as pneumococcal or D. rerio counts. Final working libraries were created after removal of two gene fractions, and clustering and functional enrichment analysis were performed.

(E) Principal-component (PC) analysis plot of pathogen transcriptional response to infection showed that the replicates cluster closely together.

(F) PC analysis plot of host response showed similar clustering behavior.

Acknowledgments

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