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. 2020 Dec 28;118(1):e1917623117. doi: 10.1073/pnas.1917623117

Fig. 3.

Fig. 3.

PACAP is a multimodal HDP that kills bacteria and fungi via synergistic mechanisms. Mechanistic fingerprinting experiments were conducted using single-cell flow cytometry. SDS (a membrane detergent), PACAP or comparator HDP LL-37, or kinocidin congeners RP-1 and ɣ-RP-1 were incubated with HDP-sensitive or HDP-resistant strains of S. aureus (A–D), S. typhimurium (E–H), or C. albicans (I–L) in the presence of specific fluorescent reporters (Materials and Methods). Six mechanistic parameters were measured in distinct pH conditions mirroring bloodstream (pH 7.5) or abscess/phagolysosomal (pH 5.5) contexts: forward scatter (FSC; cell size), side scatter (SSC; intracellular refractivity indicative of nucleic acid condensation), cellular membrane energetics (ENR; transmembrane potential), cell membrane permeability (PRM), PS accessibility (PSA; membrane turnover indicating early-stage regulated cell death), or activation of CDP (indicative of late-stage regulated cell death) pathways.