h 151  (InvivoGen)

Journal: Cancer Immunology Research

Article Title: FAM135B Deficiency Inhibits Cytotoxic T-cell Activity in Triple-Negative Breast Cancer by Blocking the IFI16-Dependent STING Pathway

doi: 10.1158/2326-6066.CIR-25-0310

Figure Lengend Snippet: FAM135B promotes cytotoxic T-cell activity by stimulating the STING–type I IFN pathway. A, BT549 and MDA-MB-231 cells expressing an empty vector or FAM135B were treated with H-151 (5 μmol/L) and cocultured with activated T cells for 48 hours. The T cells were then analyzed using flow cytometry. Representative flow cytometry plots and statistical quantification of GZMB + CD8 + cells in CD3 + tumor-infiltrating lymphocytes are shown (mean ± SD, n = 3). B, Immunoblotting analysis of the indicated proteins in TNBC cells expressing an empty vector or FAM135B under H-151 treatment. C, BT549 cells expressing an empty vector or FAM135B were treated with H-151 (5 μmol/L) and were immunostained with an anti-IRF3 antibody. Confocal microscopy images exhibited the subcellular localization of IRF3 (scale bar, 10 μm). D, Schematic representation of mouse treatment schedules. 4T1 cells (5 × 10 5 cells per mouse) expressing an empty vector or FAM135B were injected into the mammary fat pads of female BALB/c mice. Mice were injected intraperitoneally with H-151 (750 nmol per mouse) or PBS every 2 days. Three weeks after injection, the mice were humanely euthanized. E, Resected tumors from each group are shown (scale bar, 1 cm). Tumor weight was recorded, and tumor volume was calculated (mean ± SD, n = 5). F, Flow cytometry analysis revealed the population of GZMB + CD8 + cells within CD3 + tumor-infiltrating lymphocytes of mouse TNBC tissues. Representative plots and flow cytometry quantification are shown (mean ± SD, n = 3). G, The orthotopic tumor tissues were immunostained with CD8ɑ. Representative images are shown (scale bar, 20 μm). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001; ns, P > 0.05 (not significant). p-IRF, phosphorylated IRF; p-STING, phosphorylated STING; p-TBK, phosphorylated TBK.

Article Snippet: Six days after implantation, H-151 (750 nmol per mouse, MedChemExpress) was administered intraperitoneally every 2 days, cGAMP (5 mg/kg, MedChemExpress) was administered intraperitoneally every 3 days, or anti–mouse PD-1 (200 μg per mouse, Selleckchem, A2122) was administered intraperitoneally every 3 days.

Techniques: Activity Assay, Expressing, Plasmid Preparation, Flow Cytometry, Western Blot, Confocal Microscopy, Injection

Journal: Eye and Vision

Article Title: STING deficiency alleviates scar formation after glaucoma filtration surgery by suppressing p38 MAPK-induced inflammation in mice

doi: 10.1186/s40662-026-00475-3

Figure Lengend Snippet: In vivo effects of the STING inhibitor H151 on proliferation after glaucoma filtration surgery (GFS). a Representative images from the Sham, GFS, and GFS + H151 groups. Yellow circles indicate the blebs. b Bleb survival analysis of the GFS and GFS + H151 groups using Kaplan–Meier survival curves (n = 5 eyes in each group). c Histological analysis of ocular tissues by hematoxylin and eosin (H&E) staining from the Sham, GFS, and GFS + H151 groups (n = 3 eyes in each group). The triangles indicate the conjunctiva, the stars indicate the retina. d Immunofluorescence staining of α-SMA to evaluate myofibroblast activation in the Sham, GFS, and GFS + H151 groups (n = 3 eyes in each group). The triangles indicate the conjunctiva. e Masson stained tissue sections from the Sham, GFS, and GFS + H151 groups. The triangles indicate the conjunctiva, the stars indicate the retina. f Sirius red staining of tissue sections from the Sham, GFS, and GFS + H151 groups. The triangles indicate the conjunctiva, the stars indicate the retina. g Analysis of Masson staining (n = 3 eyes in each group). h Analysis of Sirius red staining (n = 3 eyes in each group). i Double immunofluorescence staining of STING (red), F4/80 (green), and DAPI (blue) in the Sham, GFS, and GFS + H151 groups (n = 3 eyes in each group). The triangles indicate the conjunctiva, the stars indicate the retina. Arrows indicate the co-localization of STING and F4/80 in the GFS group. j Immunofluorescence staining of p-p65 to assess NF-κB activation in the Sham, GFS, and GFS + H151 groups (n = 3 eyes in each group). The triangles indicate the conjunctiva, the stars indicate the retina. Arrows indicate the p-p65 expression area. k Relative mRNA expression of TNF-α, IL-18, IL-6, and IL-1β in the Sham, GFS, and GFS + H151 groups (n = 5 eyes in each group). l Relative mRNA expression of fibronectin, CTGF, COL1A1, α-SMA, and COL3A1 in the Sham, GFS, and GFS + H151 groups (n = 5 eyes in each group). Results were expressed as mean ± SD. Statistical analysis was performed using log-rank test ( b ) and one-way ANOVA followed by Bonferroni’s post-hoc test ( g , h , k , l ). #, Welch’s ANOVA with Dunnett's T3 post-hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001. STING, stimulator of interferon genes; H151, STING inhibitor; Sham, sham-operated; α-SMA, α-smooth muscle actin; F4/80, macrophage marker; DAPI, 4′,6-diamidino-2-phenylindole; p-p65, phospho-p65; NF-κB, nuclear factor kappa-B; TNF-α, tumor necrosis factor-α; IL, interleukin; CTGF, connective tissue growth factor; COL1A1, collagen type I alpha 1; COL3A1, collagen type III alpha 1

Article Snippet: H151 (HY-112693) and SB203580 (HY-10256) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: In Vivo, Filtration, Staining, Immunofluorescence, Activation Assay, Double Immunofluorescence Staining, Expressing, Marker

Journal: Eye and Vision

Article Title: STING deficiency alleviates scar formation after glaucoma filtration surgery by suppressing p38 MAPK-induced inflammation in mice

doi: 10.1186/s40662-026-00475-3

Figure Lengend Snippet: In vitro effects of H151 on inflammation and fibrosis of Ang II-induced HTFs. a Immunofluorescence staining of HTFs to identify α-SMA after the treatment with Ang II and Ang II + H151 (n = 3 eyes in each group). b , c Wound healing assay: cells were incubated with or without H151 2 h before Ang II (1 μM) treatment for 24 h. Subsequently, the wound scratches were imaged and quantified (n = 3 eyes in each group). b Representative images of the different treatment groups at different time points after the scratch. c Relative migration rates in each group. d Relative mRNA expression of TNF-α, IL-18, IL-6, and IL-1β in control, Ang II, and Ang II + H151 groups (n = 3 eyes in each group). e Relative mRNA expression of fibronectin, CTGF, COL1A1, α-SMA, and COL3A1 in control, Ang II, and Ang II + H151 groups (n = 3 eyes in each group). f , g Representative immunoblotting images: H151 significantly inhibited Ang II-induced p-p38, α-SMA, and collagen I protein expression by western blot (n = 3 eyes in each group). Results were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s post-hoc test ( d , e , g ) and two-way ANOVA followed by Bonferroni’s post-hoc test ( c ). ns, not significant, * P < 0.05, ** P < 0.01, *** P < 0.001. H151, STING inhibitor; Ang II, angiotensin II; HTFs, human Tenon’s capsule fibroblasts; α-SMA, α-smooth muscle actin; TNF-α, tumor necrosis factor-α; IL, interleukin; CTGF, connective tissue growth factor; COL1A1, collagen type I alpha 1; COL3A1, collagen type III alpha 1; p-p38, phospho-p38; DAPI, 4′,6-diamidino-2-phenylindole

Article Snippet: H151 (HY-112693) and SB203580 (HY-10256) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: In Vitro, Immunofluorescence, Staining, Wound Healing Assay, Incubation, Migration, Expressing, Control, Western Blot