sting inhibitor h 151  (MedChemExpress)

Journal: bioRxiv

Article Title: cGAS–STING induced IFN-β acts as a dual regulator of osteoclastogenesis via direct and osteoblast-mediated mechanisms

doi: 10.64898/2026.05.09.724040

Figure Lengend Snippet: (A) Effect of STING activation by 2′3′-cGAMP (BMDMs: 5 µg/mL, RAW 264.7: 10 µg/mL) on RANKL-mediated osteoclast formation. 2’3’-cGAMP were given throughout the differentiation or for BMDMs also during late stages (days 3–5/6). Representative images of osteoclasts derived from BMDMs (left) and quantification of relative osteoclast numbers per well in BMDMs and RAW 264.7 cells (right). (B) Immunoblot analysis of NFATc1 protein levels of RAW 264.7 cells following RANKL stimulation (50 ng/mL) in the presence or absence of 10 µg/mL 2′3′-cGAMP. GAPDH served as a loading control. (C) Gene expression analysis of interferon-related, macrophage-related and osteoclast-associated genes of RAW 264.7 cells 48 h after stimulation with 50 ng/mL RANKL with or without 10 µg/mL 2′3′-cGAMP. Data are presented as ratios of +cGAMP to –cGAMP. (D) Effect of STING activation by diABZI (0.01 until 10 µg/mL) on osteoclast formation. Representative images of osteoclasts derived from BMDMs (left) and quantification of relative osteoclast numbers per well in BMDMs and RAW 264.7 cells (right). (E) Induction of the interferon-responsive gene Isg15 following STING activation with diABZI (0.01 until 10 µg/mL) in BMDMs (upper) and RAW 264.7 cells (lower). Data are normalized to the unstimulated control. (F) Effect of STING activation by diABZI (0.01 until 10 µg/mL) on RANKL-induced NFATc1 expression at mRNA and protein levels after 24 h in RAW 264.7 cells. (G) Gene expression analysis of osteoclast-associated genes 48 h after stimulation with diABZI (0.01 until 10 µg/mL) and 50 ng/mL RANKL in RAW 264.7 cells. Data are normalized to the unstimulated control. (H) Effect of STING inhibition using H-151 (RAW 264.7: 40 or 400 ng/mL in DMSO, BMDMs: 400 ng/mL in DMSO) on osteoclast formation. Left and middle: quantification of relative osteoclast numbers per well upon continuous inhibitor treatment. Right: time-dependent effects of STING inhibition with inhibitor added during early stages (first 3 days) or late stages (days 3–5/6) of differentiation. BMDMs were cultured in the presence of 25 ng/mL recombinant mouse M-CSF throughout all experiments. Osteoclast numbers per well are shown relatively to the RANKL control. Heatmaps display mean values, and bar graphs show mean ± SEM with individual data points. Statistical analysis was performed using one-way ANOVA with Bonferroni post hoc test (n = 3). RL: RANKL.

Article Snippet: Where indicated, cells were treated with: cGAS agonist G3-YSD (RAW 264.7: 500 ng/mL; BMDMs: 250 ng/mL) complexed with LyoVecTM (1:100, 15 min pre-incubation), cGAS inhibitor RU.521 (10 μg/mL in DMSO) added 3 h prior to stimulation; STING agonists 2′3′-cGAMP (RAW: 10 μg/mL; BMDMs: 5 μg/mL) or diABZI (0.01–10 μg/mL), STING inhibitor H-151 (40 or 400 ng/mL in DMSO) added 2 h prior to stimulation (all InvivoGen, USA).

Techniques: Activation Assay, Derivative Assay, Western Blot, Control, Gene Expression, Expressing, Inhibition, Cell Culture, Recombinant