<p>The <i>sph</i> deletion mutant attenuates ferroptosis-associated phenotypes and bacterial virulence.</p>

<p><b>(A)</b> Growth curves of <i>S. aureus</i> WT, Δ<i>sph</i>, and Δ<i>sph</i>::<i>sph</i> strains (n = 3). <b>(B)</b> Biofilm formation of <i>S. aureus</i> WT, Δ<i>sph</i>, and Δ<i>sph</i>::<i>sph</i> strains (n = 3). <b>(C)</b> LDH release from MAC-T cells infected with <i>S. aureus</i> WT, Δ<i>sph</i>, and Δ<i>sph</i>::<i>sph</i> strains (n = 5). <b>(D)</b> Survival rates of <i>Galleria mellonella</i> infected with <i>S. aureus</i> WT, Δ<i>sph</i>, and Δ<i>sph</i>::<i>sph</i> strains (n = 9). <b>(E, F)</b> Representative immunoblot and quantitative analysis of GPX4 expression in MAC-T cells infected with <i>S. aureus</i> WT, Δ<i>sph</i>, and Δ<i>sph</i>::<i>sph</i> strains for 6 h (n = 3). <b>(G-J)</b> Representative fluorescence images and (K-N) corresponding quantitative analyses of intracellular ROS (DCFH-DA; <b>G</b>, <b>K)</b>, Fe²⁺ accumulation <b>(H, L)</b>, lipid peroxidation (C11 BODIPY 581/591; <b>I</b>, <b>M)</b>, and mitochondrial membrane potential (JC-10; J, N) in MAC-T cells infected with the indicated <i>S. aureus</i> strains (n = 3).

Scale bars, 200 μm or 100 μm. Data are presented as mean ± SD. *<i>P</i> < 0.05; **<i>P</i> < 0.01; ns, not significant.</p>