Journal: Translational Cancer Research

Article Title: Pan-cancer analysis identifies FAM111B as a biomarker for immune suppression microenvironment in low-grade gliomas

doi: 10.21037/tcr-2025-1762

Figure Lengend Snippet: FAM111B promotes the malignant progression of LGG through immunosuppression. (A) The protein expression of AKT, p-AKT, P53 and CD276 in FAM111B knockdown or overexpression LN229 and A172 cells were detected by Western blot analysis. (B) Representative bioluminescence images of FAM111B overexpression LN229 and control cells derived xenografts (n=6). (C,D) IL-10 cytokines concentration of supernatant in THP-1 cells and LN229 or A172 co-culture system, detected by ELISA. (E) Immunofluorescence for CD163 and iNOS expression to detect macrophages in glioma tissue from control and FAM111B overexpression mice. Two-tailed Student’s test, *, P<0.05; **, P<0.01; ***, P<0.001. DAPI, 4',6-diamidino-2-phenylindole; ELISA, enzyme linked immunosorbent assay; FAM111B, FAM111 trypsin-like peptidase B; IL-10, interleukin-10; iNOS, inducible nitric oxide synthase; LGG, lower-grade glioma; NC, negative control; OE-FAM111B, overexpression of FAM111B; siFAM111B, small interfering RNA targeting FAM111B; THP-1, tumor-derived human promonocytic cell line-1; WT, wild-type.

Article Snippet: Subsequently, supernatant from THP-1 cells was collected, and the levels of interleukin-10 (IL-10) were assessed using an ELISA kit (Elabscience) to evaluate the polarization degree of macrophages.

Techniques: Expressing, Knockdown, Over Expression, Western Blot, Control, Derivative Assay, Concentration Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Two Tailed Test, Negative Control, Small Interfering RNA

Journal: Frontiers in Immunology

Article Title: Carbohydrate fatty acid monosulphate ester adjuvant enhances the immunogenicity of influenza antigens via TLR4/2-dependent mechanisms

doi: 10.3389/fimmu.2026.1787181

Figure Lengend Snippet: CMS activates DCs through TLR4 and TLR2-dependent mechanisms. (A) Heat map representation depicting the expression of different TLRs comparing unstimulated DCs vs CMS-stimulated DCs. (B) THP-1 human monocytic cell line stably expressing a secreted embryonic alkaline phosphatase (SEAP) reporter inducible by NF-kB was treated for 24 hrs with CMS at the indicated concentrations. The activation of NF-κB was assessed by measuring the activity of SEAP in the supernatant using QUANTI−Blue™ Solution. Results of THP1 MD2-CD14-TLR4, THP1 and THP1 KO-TLR4 are shown (mean ± SD, n=4). (C, E) Effect of CLI095 (TLR4 pathway inhibitor), or TL2-C29 (TLR2 pathway inhibitor), compared to DMSO (solvent control) on the expression of CMS-induced DC maturation markers. Scatter plots representing the mean ± SD (n = 8 donors) values of CD80, CD86, and HLA-DR are presented. (D, F) Inhibitory effect of CLI095 or TL2-C29 on the secretion (mean ± SD, n = 8) of IL-8, IL-6, and IL-12p70 (all in pg/ml) by CMS-stimulated DCs. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 as analyzed by one-way ANOVA with Tukey’s multiple comparisons post-test (C-F) . CA, cells alone; DMSO, dimethyl sulfoxide; SD, standard deviation.

Article Snippet: THP1-Dual, THP1-Dual MD2-CD14-TLR4, and THP1-Dual KO-TLR4 reporter cells (Invivogen) were resuspended at a concentration of 100,000 cells per well in a 96-well U-bottom plate in 200 μl RPMI 1640 Medium, GlutaMAX Supplement (Gibco), supplemented with 25mM HEPES, 10% FBS, 100 IU/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Expressing, Stable Transfection, Activation Assay, Activity Assay, Solvent, Control, Standard Deviation