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. Author manuscript; available in PMC: 2023 Oct 19.
Published in final edited form as: Cell Rep. 2023 Aug 22;42(9):113024. doi: 10.1016/j.celrep.2023.113024

Figure 2. SE isogenic mutant strains reveal specific PSMs drive keratinocyte inflammation.

Figure 2.

(A) PCR confirmation gel for specific SE knockout strains generated for this study. SE hld point mutation in start codon for gene confirmed through DNA sequencing and indicated by a +/−. For knockout strain growth curves, see also Figures S1C and S1D.

(B) qPCR analysis of up-regulated inflammatory markers in keratinocytes treated for 3 h with 10% sterile-filtered conditioned medium of overnight growths of SE wild-type (WT) and specific knockout strains (n = 4).

(C and D) (C) hIL-8 ELISA and (D) LDH release analysis from conditioned medium of keratinocytes treated for 24 h with 10% sterile-filtered conditioned medium of overnight growths of SE wild-type (WT) and specific knockout strains (n = 4).

(E–H) Mass spectrometric validation of the presence or absence of PSMs in spent media for S. epidermidis 1457 WT and knockout strains. Results are averages of selected ion chromatogram peak areas for injections of biological triplicates of S. epidermidis spent media (TSB). The ions were detected in the quadropoly protonated form, [M+4H]4+. Results are representative of at least two independent experiments. Mean ± SEM and a parametric unpaired one-way ANOVA analysis was used to determine statistical significance: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. See also Figure S2.