Journal: Nature Communications

Article Title: CD133 + PD-L1 + cancer cells confer resistance to adoptively transferred engineered macrophage-based therapy in melanoma

doi: 10.1038/s41467-025-55876-0

Figure Lengend Snippet: a Expression of PD-L1 in B16F10 cancer cells or NIH3T3 fibroblast cells in the co-culture with or without TGF-β. The expression of PD-L1 was normalized with the corresponding untreated B16F10 or NIH3T3 cells cultured alone. Two-way ANOVA with Sidak’s post-test ( n = 4 biologically independent samples). b Expression of PD-L1 in the cancer cells and CAFs of mice following different treatments, normalized with the group of untreated day 8. Untreated day 16, the mice without treatment at day 16 reached a similar tumor size in volume ( ~ 500 mm 3 ) to that of mice following the NPs-MΦ 1 treatment at day 35. One-way ANOVA with Tukey’s post-test ( n = 6 biologically independent samples for untreated day 8, n = 5 biologically independent samples for untreated day 16 or NPs-MΦ 1 day 35). c Identification of CD133 + PD-L1 + from the B16-GFP cancer cells of mice receiving the adoptive NPs-MΦ therapy. Tumors were collected from the NPs-MΦ 1 -treated mice at day 35. One-way ANOVA with Tukey’s post-test ( n = 5 biologically independent mice). Right, representative dot plots of the flow cytometric analysis. d Flow cytometric analysis of TGF-β, IL-10 or VEGF-α production in GFP + CD133 - PD-L1 - , GFP + CD133 + PD-L1 - , GFP + CD133 - PD-L1 + and GFP + CD133 + PD-L1 + cancer cells of mice following the NPs-MΦ 1 treatment at day 35, respectively. The expression of TGF-β, IL-10, or VEGF-α were normalized with the corresponding GFP + CD133 + PD-L1 - cancer cells. One-way ANOVA with Tukey’s post-test ( n = 5 biologically independent samples for GFP + CD133 - PD-L1 - cancer cells, n = 4 biologically independent samples for GFP + CD133 + PD-L1 - , GFP + CD133 - PD-L1 + or GFP + CD133 + PD-L1 + cancer cells). e Volcano plot indicating differentially expressed genes relevant to the TGF-β pathway. The x- axis indicates the log 2 -fold change for GFP + CD133 + PD-L1 + versus GFP + CD133 - PD-L1 - cancer cells. The y- axis indicates the −1 × log 10 of the Q value. Statistical significance was calculated using the two-sided Wald test for DESeq2 data. Source data are provided as a Source Data file.

Article Snippet: Mouse B16F10 melanoma cell line (cat. CRL-6475), mouse NIH3T3 embryonic fibroblast cell line (cat. CRL-1658) and human A375 melanoma cell line (cat. CRL-1619) were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Co-Culture Assay, Cell Culture

Journal: bioRxiv

Article Title: In vitro -generated inflammatory fibroblasts secrete extracellular matrix with biochemical and biophysical properties similar to tissue-remodelling fibroblasts

doi: 10.1101/2024.09.26.614950

Figure Lengend Snippet: A) Schematic representation of the stimuli used to induce fibroblast differentiation in vitro in 3D. NIH3T3 fibroblasts were mixed with a drop of rat tail fibrillar collagen I (1.7mg/mL polymerized at 37°C) and cultured for 4 days in the presence of medium alone or medium supplemented with the indicated stimuli. B-C) Log2 gene expression of myCAF markers Acta2 and Tagln (B) and Ccl2, Il6 and Lif (C) of in vitro -induced subtypes normalized to uninduced. Housekeeping genes are Ppia and B2m . Data were analysed by one-way ANOVA followed by Tukey’s multiple comparisons test of three independent experiments. *p<0.05; **p<0.01. D) t-SNE plots of 3,392 stromal fibroblasts of interest from CRC dataset (top) and PDAC dataset (bottom), color-coded by sub-cell type and sample origin, and 6,781 stromal fibroblasts of interest. E) Dot-plots representing average gene expression, and percentage of cells expressing Acta2, Tagln, Il6, Ccl2 and Lif . From the CRC datasets (top), a comparison is made between subtypes Myofibroblasts, Stromal 1, Stromal 2, and Stromal 3 of tumor origin. From the PDAC dataset (bottom), myCAFs and iCAFs are compared.

Article Snippet: Embryonic mouse fibroblast cell line NIH3T3 was obtained from ATCC (CRL-1658) and cultured in high-glucose DMEM with 4.5 g/L glucose (Gibco, #10938025) supplemented with 10% fetal calf serum (FCS) (Cytiva, SH30541.03), penicillin-streptomycin (Sigma-Aldrich, #P4333), 200 mM L-glutamine (Gibco, #25030-024) and 1% sodium pyruvate (Gibco, #25030-024), namely complete medium.

Techniques: In Vitro, Cell Culture, Gene Expression, Expressing, Comparison