Journal: Nature Microbiology

Article Title: Skin androgens regulate Staphylococcus aureus pathogenicity via quorum sensing

doi: 10.1038/s41564-026-02261-2

Figure Lengend Snippet: a , The S. aureus HG003 strain was treated with 10 nM testosterone or vehicle to early-log phase (OD 600 = 0.4). Volcano plot showing genes with >4-fold expression change (green) by RNA sequencing. b , Schematic of S. aureus agr quorum-sensing system. c , agr -induced bioluminescence of HG003 agr- P3:: lux quorum-sensing reporter in vivo treated with 10 nM testosterone, DHT, AIP-I or vehicle ( n = 3). Means ± s.e.m. (error bars) are plotted. # P < 0.0001 by 2-way ANOVA compared with vehicle. d , Quantitative real-time PCR (qRT-PCR) of S. aureus HG003 (type-I strain) treated with 10 nM testosterone, AIP-I or vehicle ( n = 3) until mid-exponential growth (OD 600 = 0.6). Gene expression is normalized to gyrA . Means ± s.e.m. (error bars) are plotted. ** P < 0.01, *** P < 0.001 by unpaired 2-tailed Student’s t- test. e , f , qRT-PCR of MRSA type-II (USA100) ( e ) and MRSA type-III (MW2) ( f ) strains cultured to mid-log phase, treated with 10 nM testosterone, AIP-II or AIP-III ( n = 3). psm α expression is normalized to gyrA . Means ± s.e.m. (error bars) are plotted. * P < 0.05 by unpaired 2-tailed Student’s t- test. g , qRT-PCR for psmα in S. aureus isolates from patients with AD treated with 10 nM testosterone or vehicle ( n = 3) to mid-exponential growth. Means ± s.e.m. (error bars) are plotted. * P < 0.05, ** P < 0.01 by unpaired 2-tailed Student’s t- test. h , Percentage of S. aureus -induced RBC haemolysis with and without 10 nM testosterone, DHT or AIP-I ( n = 4 human donors, 2 men and 2 women). Means ± s.e.m. (error bars) are plotted. **** P < 0.0001, by 1-way ANOVA. i , Percentage of S. aureus -induced skin cell cytotoxicity with and without 10 nM testosterone, AIP-I or AIP-I and testosterone ( n = 3). Means ± s.e.m. (error bars) are plotted. ** P < 0.01, *** P < 0.001 by 1-way ANOVA. j , agr -induced bioluminescence in vivo. Left, epicutaneous infection of male and female Hsd3b6 fl/fl ( n = 7 males and n = 5 females) and Hsd3b6 ∆skin mice ( n = 6 males and n = 5 females) with bioluminescent CA-MRSA- agr -P3:: lux with agr -induced bioluminescence quantified over time. Right, representative bioluminescence images. $ P < 0.0001, ** P < 0.01 and non-significant (NS) by 2-way ANOVA with comparisons. Aggregate of two experiments. Means ± s.e.m. (error bars) are plotted. See Extended Data Figs. – . Panel b created in BioRender; Harris, T. https://biorender.com/wd041bk (2025).

Article Snippet: Experiments were completed in triplicate, with the agr -type-specific AIPs AIP-I (catalogue number 4515-v; Peptide Institute), AIP-II (catalogue number 4516-v; Peptide Institute) and AIP-III (catalogue number 4517-v; Peptide Institute) as positive control.

Techniques: Expressing, RNA Sequencing, In Vivo, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Gene Expression, Cell Culture, Infection

Journal: Nature Microbiology

Article Title: Skin androgens regulate Staphylococcus aureus pathogenicity via quorum sensing

doi: 10.1038/s41564-026-02261-2

Figure Lengend Snippet: a – c , S. aureus biosynthetic mutant (HG003-Δ agrBD ) and its respective agr- P3 bioluminescent reporter strain (Δ agrBD agr- P3 :: lux ) were treated with 10 nM testosterone, AIP-I or untreated. a , agr -induced bioluminescence of agr- P3:: lux was recorded every hour using a plate reader ( n = 12). Statistics of testosterone and AIP-I compared with the vehicle. Means ± s.e.m. (error bars) are plotted. # P < 0.0001 by 2-way ANOVA compared with vehicle. b , Percentage of bacterially induced haemolysis and skin cell cytotoxicity ( n = 3 replicates of RBCs from a single donor). c , qRT-PCR of psmα expression in the biosynthetic mutant strain treated with 10 nM testosterone, AIP-I and vehicle ( n = 3), normalized to gyrA expression. d , qRT-PCR for psmα expression in the biosynthetic mutant strain treated with AIP-I and 10 nM or 100 nM of testosterone ( n = 3). e , f , qRT-PCR for psmα expression in the biosynthetic mutant strain treated with AIP-II ( e ) or AIP-III ( f ) alone or in combination with increasing concentrations of testosterone (10 nM to 10 µM) ( n = 3). b – f , Means ± s.e.m. (error bars) are plotted. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by 1-way ANOVA. g , h , Male Hsd3b6 fl/fl ( n = 8) and Hsd3b6 ∆skin ( n = 5 per group) mice were epicutaneously infected with 1 × 10 6 CFU of the biosynthetic mutant strain constitutive reporter (Δ agrBD :: lux ) treated topically with testosterone, AIP-I or untreated, with bioluminescence quantified over time ( g ) and representative bioluminescence images ( h ). Results are an aggregate of two experiments. Means ± s.e.m. (error bars) are plotted. # P < 0.01 by 2-way ANOVA of Hsd3b6 fl/fl compared with vehicle-treated Hsd3b6 ∆skin mice. & P < 0.01 by 2-sided Mann–Whitney U- test of vehicle compared with testosterone-treated Hsd3b6 ∆skin mice on day 1. See Extended Data Fig. .

Article Snippet: Experiments were completed in triplicate, with the agr -type-specific AIPs AIP-I (catalogue number 4515-v; Peptide Institute), AIP-II (catalogue number 4516-v; Peptide Institute) and AIP-III (catalogue number 4517-v; Peptide Institute) as positive control.

Techniques: Mutagenesis, Quantitative RT-PCR, Expressing, Infection, MANN-WHITNEY

Journal: Nature Microbiology

Article Title: Skin androgens regulate Staphylococcus aureus pathogenicity via quorum sensing

doi: 10.1038/s41564-026-02261-2

Figure Lengend Snippet: a , Biosynthetic mutant Δ agrBD treated with increasing concentrations of testosterone. RNAIII , agrA , agrC , and psmα quantified by qRT-PCR ( n = 3).Means ± SEM (error bars) are plotted. * p < 0.05; *** p < 0.001, ns, not significant by one-way ANOVA with Tukey’s post-hoc test (two-sided), adjusted for multiple comparisons. b , Growth curves of Δ agrBD treated with 10 nM testosterone, DHT or vehicle ( n = 3), no significant differences from vehicle by two-way ANOVA. Means ± SEM (error bars) are plotted. c , Δ agrBD::lux treated with 10 nM testosterone, DHT, progesterone, or estradiol ( n = 3). Bioluminescence was measured. Means ± SEM (error bars) are plotted. d , RNAIII expression in Δ agrBD treated with 10 nM AIP-I and 10 or 100 nM testosterone ( n = 3) was quantified. e , Neutrophil killing induced by Δ agrBD treated with 10 nM testosterone, AIP-I, or vehicle was measured ( n = 3 from a single donor). Means ± SEM are plotted. ( c - e ) * p < 0.05; ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant by one-way ANOVA (two-sided). ( c ) # p < 0.0001 and ns by two-way ANOVA (two-sided). versus estradiol. f , Schematic showing inhibition of AgrC signaling by non-cognate AIPs. g,h , Δ agrBD strains were treated with AIP-II ( g ) or AIP-III ( h ) alone or with increasing concentrations of testosterone (10nM–10 μM). RNAIII , agrA , and agrC were quantified ( n = 3). Means ± SEM (error bars) are plotted. * p < 0.05, ** p < 0.01; *** p < 0.001, **** p < 0.0001, ns by one-way ANOVA. i , Growth curve of Δ agrC treated with 10 nM testosterone showed no significant difference from vehicle ( n = 3) by two-way ANOVA. Means ± SEM (error bars) are plotted. j,k , Neutrophil killing ( n = 3 replicates of RBCs from a single donor) ( j ) and skin cell cytotoxicity ( n = 3) (k) analysed in Δ agrC treated with 10 nM testosterone, DHT, AIP-I, or combinations. ns by one-way ANOVA. Means ± SEM (error bars) are plotted. l,m , Δ agrC ( l ) and Δ agrA (m) mutants were treated with 10 nM testosterone, AIP-I, or vehicle, ( n = 3) and psmα and RNAIII were quantified. Means ± SEM (error bars) are plotted. ns, not significant by one-way ANOVA. n , Δ agrC mutant strain complemented (Comp) with AgrC on a plasmid treated with 10 nM testosterone, AIP-I, or vehicle ( n = 3) and agrA and RNAIII were quantified. Means ± SEM (error bars) are plotted. * p < 0.05 by one-way ANOVA. Panel f created in BioRender; Harris, T. https://biorender.com/dzfdpzh (2025).

Article Snippet: Experiments were completed in triplicate, with the agr -type-specific AIPs AIP-I (catalogue number 4515-v; Peptide Institute), AIP-II (catalogue number 4516-v; Peptide Institute) and AIP-III (catalogue number 4517-v; Peptide Institute) as positive control.

Techniques: Mutagenesis, Quantitative RT-PCR, Expressing, Inhibition, Plasmid Preparation