Fig 6 - <p> </p>

<p><i>S</i><i>. aureus</i> Sph activates STING to promote autophagy-lysosome-dependent GPX4 degradation and ferroptosis. <b>(A)</b> Expression of GPX4 protein in HC11 cells treated with 32 μg/mL recombinant Sph and C-176 (5 μM) for 6 h. <b>(B)</b> Expression of of LC3B-I and LC3B-II levels in HC11 cells treated with recombinant Sph (32 μg/mL) for 6 h. <b>(C)</b> GPX4 expression in HC11 cells treated with recombinant Sph (32 μg/mL) alone or in combination with NH₄Cl (10 mM) or chloroquine (20 μM) for 6 h. <b>(D)</b> Expression of GPX4 protein in HC11 cells treated with 32 μg/mL recombinant Sph alone or in combination with 3-MA (5 mM) or bafilomycin A1 (100 nM) for 6 h. <b>(E-G)</b> Representative confocal images and corresponding colocalization analyses of STING with GPX4 <b>(E)</b>, LC3B with GPX4 <b>(F)</b>, and LAMP2B with GPX4 <b>(G)</b> in HC11 cells treated with recombinant Sph (32 μg/mL), cGAMP (2 μg/mL), or C-176 (5 μM), as indicated, for 6 h.

Scale bars, 10 μm. <b>(H, I)</b> Reciprocal co-immunoprecipitation analysis of Flag-GPX4 and Myc-STING in co-transfected NIH/3T3 cells using anti-Flag (H) or anti-Myc <b>(I)</b> antibodies. <b>(J)</b> Ratiometric imaging of Golgi luminal pH in NIH/3T3 cells expressing MGAT2-RpHLuorin2 following treatment with cGAMP (2 μg/mL) or bafilomycin A1 (100 nM). The 405/488 nm fluorescence ratio was used as an indicator of Golgi pH (MGAT2-RpHLuorin2).