sting inhibitor c 176  (MedChemExpress)

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Heme aggravates bacterial infection induced STING activation . ( A, B ) Western blot analysis and quantification of p-STING/STING, p-TBK1/TBK1, and p-IRF3/IRF3 in HUVECs treated with heme (10LµM), heat-killed E. coli (MOI:10), or both for 6 hours (n=6/group). ( C ) Structural modeling of heme (pink) binding to the STING dimer complex using AlphaFold. ( D, E ) Native gel Western blot analysis of STING oligomerization in HUVECs treated with heme, E. coli , cGAMP, or their combinations. ( F, G ) Immunofluorescence staining of STING aggregates in HUVECs transfected with STING-HA and TBK1-Flag plasmids, followed by treatment with heme, E. coli , cGAMP, or their combinations. Scale bars, 20Lμm. ( H, I ) Western blot analysis and quantification of p-STING/STING, p-TBK1/TBK1, and p-IRF3/IRF3 in heart tissues from sham and septic mice treated with heme (n=4/group). All data are presented as mean ± SD. Statistical significance was determined using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: For treatment experiments, HUVECs were exposed to the following stimuli and inhibitors, either alone or in combination as indicated: heme (10 μM, Hemin, Sigma, Cat: 51280-5G), heat-killed E. coli (MOI: 20, ATCC, Cat: BAA197), STING inhibitor (2.5µM, C-176, MCE, Cat: HY-112906).

Techniques: Infection, Activation Assay, Western Blot, Binding Assay, Immunofluorescence, Staining, Transfection, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: Heme aggravates bacterial infection induced STING activation . ( A-D ) Co-immunoprecipitation (Co-IP) assay indicate interaction between TBK1 and STING in HUVECs transfected with STING-HA and TBK1-Flag plasmids, followed by treatment with heme (10LµM), heat-killed E. coli (MOI:10), cGAMP (5Lµg/mL), or their combinations for 1 hour. ( E, F ) Native PAGE Western blot analysis demonstrating STING oligomerization in HUVECs transfected with STING-HA and TBK1-Flag plasmids under the same treatment conditions. ( G, H ) Immunofluorescence staining of STING aggregates in HUVECs following treatment with heme, E. coli , cGAMP, or combinations. Scale bars, 20Lμm. All data are presented as mean ± SD. Statistical significance was determined using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: For treatment experiments, HUVECs were exposed to the following stimuli and inhibitors, either alone or in combination as indicated: heme (10 μM, Hemin, Sigma, Cat: 51280-5G), heat-killed E. coli (MOI: 20, ATCC, Cat: BAA197), STING inhibitor (2.5µM, C-176, MCE, Cat: HY-112906).

Techniques: Infection, Activation Assay, Co-Immunoprecipitation Assay, Transfection, Clear Native PAGE, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: STING activation contributes to sepsis induced cardiac endothelial senescence . ( A, B ) Co-immunofluorescent staining and quantification of the senescence marker p21 with the endothelial cell marker CD31 in cardiac tissues from sham and septic mice treated with a STING inhibitor or vehicle control at 24 hours post-CLP (n=6 mice/group). Scale bars, 50Lμm. ( C ) Representative echocardiographic images assessing ejection fraction (EF) and fractional shortening (FS) at 24 hours, 7 days, and 14 days post-CLP in mice treated with vehicle or STING inhibitor. ( D ) Western blot analysis of p-STING/STING, p-TBK1/TBK1, and p-IRF3/IRF3 in HUVECs treated with heme + heat-killed E. coli , with or without the STING inhibitor C-176 (2.5LμM) (n=6/group). ( E, F ) Echocardiographic assessment of EF and FS in septic mice treated with STING inhibitor or vehicle control at 24 hours, 7 days, and 14 days post-CLP, as shown in (C) (n=6 mice/group). ( G ) Quantification of Western blot analysis of p-STING/STING, p-TBK1/TBK1, and p-IRF3/IRF3 in the indicated HUVEC groups, as shown in ( D ) (n=4/group). ( H, I ) Western blot analysis and quantification of senescence markers p16, p21, and acetylated p53 in the indicated HUVEC groups (n=6/group). ( J, K ) β-galactosidase (β-gal) staining and quantification of HUVECs treated with combined heme and E. coli with or without the STING inhibitor C-176 (n=6/group). Scale bars, 100Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: For treatment experiments, HUVECs were exposed to the following stimuli and inhibitors, either alone or in combination as indicated: heme (10 μM, Hemin, Sigma, Cat: 51280-5G), heat-killed E. coli (MOI: 20, ATCC, Cat: BAA197), STING inhibitor (2.5µM, C-176, MCE, Cat: HY-112906).

Techniques: Activation Assay, Staining, Marker, Control, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: STING inhibition attenuates sepsis-induced cardiac endothelial senescence . ( A, B ) Immunofluorescent co-staining and quantification of the senescence marker p16 with the endothelial marker CD31 in heart tissues from sham and septic mice treated with a STING inhibitor or vehicle at 24 hours post-CLP (n=5-6 mice/group). Scale bars, 50Lμm. ( C–E ) Immunofluorescent co-staining and quantification of senescence markers p16 or p21 with the endothelial marker CD31 in heart tissues from sham and septic mice treated with a STING inhibitor or vehicle at 14 days post-CLP (n=4-6 mice/group). Scale bars, 50Lμm. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: **PL

Article Snippet: For treatment experiments, HUVECs were exposed to the following stimuli and inhibitors, either alone or in combination as indicated: heme (10 μM, Hemin, Sigma, Cat: 51280-5G), heat-killed E. coli (MOI: 20, ATCC, Cat: BAA197), STING inhibitor (2.5µM, C-176, MCE, Cat: HY-112906).

Techniques: Inhibition, Staining, Marker, Two Tailed Test

Journal: bioRxiv

Article Title: Heme drives cardiac endothelial senescence in sepsis via STING activation

doi: 10.1101/2025.06.18.660159

Figure Lengend Snippet: STING inhibition restores endothelial reparative function in vitro . ( A, B ) β-galactosidase staining and quantification of HUVECs treated with septic plasma, with or without STING inhibitor (C-176, 2.5LµM). ( C, D ) Immunofluorescent staining and quantification of senescence marker p21 in HUVECs treated with septic plasma, with or without STING inhibitor (n=6/group). Scale bars, 50Lμm. ( E–H ) Endothelial proliferation (BrdU) and tube formation assays following treatment with combined heme and E. coli, with or without STING inhibitor. All data are presented as mean ± SD. Statistical analysis was performed using the unpaired two-tailed Student’s t-test: *PL

Article Snippet: For treatment experiments, HUVECs were exposed to the following stimuli and inhibitors, either alone or in combination as indicated: heme (10 μM, Hemin, Sigma, Cat: 51280-5G), heat-killed E. coli (MOI: 20, ATCC, Cat: BAA197), STING inhibitor (2.5µM, C-176, MCE, Cat: HY-112906).

Techniques: Inhibition, In Vitro, Staining, Clinical Proteomics, Marker, Two Tailed Test