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: Biomedicines

Article Title: A Matrix-Free 3D Hepatocyte–Macrophage Co-Culture Spheroid Model for Dual Assessment of Lipid Accumulation and NF-κB-Mediated Inflammatory Activation Under Glucolipotoxic Stress

doi: 10.3390/biomedicines14040792

Figure Lengend Snippet: Morphology and metabolic activity of HepG2/THP-1 spheroids at different seeding densities. ( A ) Representative Live/Dead staining at Day 8 of co-culture spheroids seeded at 3000 (3k) or 6000 (6k) total cells/well. Viable cells are calcein-AM-positive (green) and dead cells are EthD-1-positive (red); corresponding nuclear counterstaining (DAPI, blue) is shown in the lower panels. Scale bars: 100 µm. ( B ) Metabolic activity during maturation (Days 3, 6, 8, and 10) measured by PrestoBlue™ fluorescence (Ex/Em 560/590 nm), normalized to the starting seeded cell number, and reported as normalized RFU. The graph shows one representative independent experiment; data are presented as mean ± SD of 8 technical replicate wells per condition. The experiment was repeated independently three times with comparable results. Two-way ANOVA (factors: time and seeding density) with Šídák’s multiple comparisons. Abbreviations: RFU, relative fluorescence units; EthD-1, ethidium homodimer-1; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: HepG2 and THP-1 NF-κB-Luc2 were authenticated by STR profiling (ATCC ® Lot number 70050519 and 70059144) and were mycoplasma-negative.

Techniques: Activity Assay, Staining, Co-Culture Assay, Fluorescence

Journal: Biomedicines

Article Title: A Matrix-Free 3D Hepatocyte–Macrophage Co-Culture Spheroid Model for Dual Assessment of Lipid Accumulation and NF-κB-Mediated Inflammatory Activation Under Glucolipotoxic Stress

doi: 10.3390/biomedicines14040792

Figure Lengend Snippet: Hepatocyte and macrophage marker expression during spheroid phenotypic remodeling. HepG2/THP-1 spheroids were harvested at Days 3, 6, 8, and 10 for bulk-spheroid RNA extraction followed by qPCR. ( A ) Hepatocyte-associated transcripts (ALB, AFP, CYP3A4) expressed as fold change versus 2D HepG2 monolayers (calibrator = 1.00). ( B ) Macrophage-associated transcripts (CD14, CD64, CD68, CD206, MARCO, TREM2) expressed as fold change versus 2D PMA-differentiated THP-1 cells (calibrator = 1.00). ( C ) Representative immunofluorescence at Day 8 showing albumin (ALB, green) and CD14 (red); nuclei counterstained with DAPI (blue). Scale bars: 100 µm. The graph shows one representative independent experiment; data are presented as mean ± SD of n = 4 biological replicates per condition, each biological replicate generated by pooling 2 spheroids. The experiment was repeated independently three times with comparable results. One-way ANOVA with Tukey’s multiple comparisons; groups not sharing letters differ significantly ( p < 0.05). Abbreviations: ALB, albumin; AFP, alpha-fetoprotein; CYP3A4, cytochrome P450 family 3 subfamily A member 4; CD, cluster of differentiation; MARCO, macrophage receptor with collagenous structure; TREM2, triggering receptor expressed on myeloid cells 2; DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: HepG2 and THP-1 NF-κB-Luc2 were authenticated by STR profiling (ATCC ® Lot number 70050519 and 70059144) and were mycoplasma-negative.

Techniques: Marker, Expressing, RNA Extraction, Immunofluorescence, Generated