a, Outline for experiments (b-d). b, qPCR detection of S. pneumoniae psaA gene expression in tibia versus skull normalized to sham (mean ± SD; n=4; P value represents a Mann-Whitney two-tailed rank test). c, Flow cytometry gating. d, Quantitation of BrdU+ lineage- Sca-1+ c-kit+ hematopoietic progenitors (mean ± SD; n=11 sham, 12 meningitis; P values represent unpaired, two-tailed t-tests). e, Experimental outline. f, IVM of skull marrow at 4–6 hours after intracisternal injection of GFP+
Streptococcus pneumoniae JWV500. Vasculature was labeled with CD31/Sca-1 and CSF with IC ovalbumin. g, inset depicts large GFP+ bacterial areas (arrows)(f and g are representative data from 2 independent experiments). h, inset shows smaller GFP+ areas, presumably bacterial colonies (n=2). i, Experimental outline. Flow-sorted LSK underwent membrane staining encoding their genotype, Myd88−/− (green) and wild type LSK (magenta) were transferred to a recipient mouse in which meningitis was induced. IVM was done 48 hours after infection. j, Intravital microcopy images show less Myd88−/− LSK compared to wild type control LSK. k, Quantification of LSK (mean ± SD; n = 7 recipient mice, P value represents an unpaired, two-tailed t-test).