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ATCC melanoma cells
Melanoma Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 5237 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/a+375+cells/A-375/us12653793-273-20-22
Average 99 stars, based on 5237 article reviews

melanoma cells - by Bioz Stars, 2026-10

99/100 stars

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Cell Culture:

Article Title: A structural and functional comparison between two recombinant human lubricin proteins: Recombinant human proteoglycan-4 (rhPRG4) vs ECF843.
Article Snippet: Proteoglycan 4 (PRG4, lubricin) is a mucin-like glycoprotein present on the ocular surface that has both boundary lubricating and anti-inflammatory properties.. Full-length recombinant human PRG4 (rhPRG4) has been shown to be clinically effective in improving signs and symptoms of dry eye disease (DED).. In vitro, rhPRG4 has been shown to reduce inflammation-induced cytokine production and NFκB activity in corneal epithelial cells, as well as to bind to and inhibit MMP-9 activity.

Article Title: Dual Checkpoint Aptamer Immunotherapy: Unveiling Tailored Cancer Treatment Targeting CTLA-4 and NKG2A
Article Snippet: HCT-15, COLO-205, LNCaP clone FGC, BT-549 cells, and human peripheral blood mononuclear cells (hPBMCs) were cultured in RPMI-1640 medium (ATCC, Manassas, VA, USA). .. A-375 cells were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) (Gibco, Grand Island, NY, USA), while A-549 cells were cultured in F-12K medium (ATCC, Manassas, VA, USA). .. The BT-474 cell line was cultured in Hybri-Care medium (ATCC, Manassas, VA, USA) supplemented with 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA).

Modification:

Article Title: Solution chemical properties and anticancer potential of 8-hydroxyquinoline hydrazones and their oxidovanadium(IV) complexes.
Article Snippet: The experiments were carried out with two replicates, and band intensities were further quantified and analysed using ImageJ software. .. A-549 (ATCC, CCL-185) cells were grown in Dulbecco’s Modified Eagle Medium F-12 (Gibco #11320033) and A-375 cells (ATCC, CRL3222) were grown in Dulbecco’s Modified Eagle Medium with high glucose (Gibco #11965118), supplemented with 10% FBS (Gibco, 10500064) and penicillin-streptomycin (10,000 U/mL, Gibco, 15140122). ..

other:

Article Title: Ultra-long stable biomimetic nanoparticle Click-ed-to-cancer membrane for anti-cancer treatment
Article Snippet: Upon sub-cellular fractionation, membranes of (i) H1573 cells (lung adenocarcinoma) encapsulated gold nanoparticles incubated in RPMI 1640 Medium (R8758-500 ML), 10% FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); (ii) OVCA432 cells (high-grade serous epithelial ovarian cancer) encapsulated gold nanoparticles incubated in Medium 199 (M4530-500ML, Merc Life Science Pty Ltd), MCDB 105 Medium (M6395-1L, Merc Life Science Pty Ltd), 1% Penicillin/Streptomycin solution (G255, abm); (iii) CaCO2 [HTB-37, ATCC] (Colorectal Cancer) encapsulated gold nanoparticles incubated in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco); (iv) UM-HACC-2A cells (adenoid cystic carcinoma) (T8326, abm) encapsulated gold nanoparticles incubated in optimized salivary gland medium consisted of PriGroIII (TM003, abm), 10% fetal bovine serum (A56695-01, gibco), 2 mM L-glutamine (G275, abm), 0.4 μg/mL hydrocortisone (H0135-1MG, Sigma Aldrich), 20 ng/mL recombinant human epidermal growth factor (Z100139. abm), 5 μg/mL recombinant human insulin (Z101065, abm), 1% Penicillin/Streptomycin solution (G255, abm); (v) A-375 cells (malignant melanoma) (CRL-1619, ATCC) encapsulated gold nanoparticles incubated in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); (vi) SCC9 cells (oral cancer) encapsulated gold nanoparticles incubated in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); (vii) CW1474 cell line (CRL-3529, ATCC) (esophageal adenocarcinoma) encapsulated gold nanoparticles incubated in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); and (viii) MCF7 [HTB-22, ATCC] (Breast Cancer) encapsulated gold nanoparticles incubated in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); at 4 °C for 12-months.

Article Title: Ultra-long stable biomimetic nanoparticle Click-ed-to-cancer membrane for anti-cancer treatment
Article Snippet: H1573 cells (lung adenocarcinoma) (CRL-5877, ATCC) cultured in RPMI 1640 Medium (R8758-500 ML), 10% FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); OVCA432 cells (high-grade serous epithelial ovarian cancer) (cells kindly gifted by Dr Charlie Ahn, Macquarie University) cultured in Medium 199 (M4530-500ML, Merc Life Science Pty Ltd), MCDB 105 Medium (M6395-1L, Merc Life Science Pty Ltd), 1% Penicillin/Streptomycin solution (G255, abm); CaCO2 [HTB-37, ATCC] (Colorectal Cancer) cultured in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco); UM-HACC-2A cells (adenoid cystic carcinoma) (T8326, abm) cultured in optimized salivary gland medium consisted of PriGroIII (TM003, abm), 10% fetal bovine serum (A56695-01, gibco), 2 mM L-glutamine (G275, abm), 0.4 μg/mL hydrocortisone (H0135-1MG, Sigma Aldrich), 20 ng/mL recombinant human epidermal growth factor (Z100139. abm), 5 μg/mL recombinant human insulin (Z101065, abm), 1% Penicillin/Streptomycin solution (G255, abm); A-375 cells (malignant melanoma) (CRL-1619, ATCC) cultured in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); SCC9 cells (oral cancer) (cells kindly gifted by Dr Charbel Darido, University of Melbourne) cultured in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); CW1474 cell line (CRL-3529, ATCC) (esophageal adenocarcinoma) cultured in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm); and MCF7 [HTB-22, ATCC] (Breast Cancer) cultured in High-glucose Dulbecco’s modification of Eagle medium (DMEM) (D5796-500ML, Thermo Fisher Scientific), FBS (A56695-01, gibco), 1% Penicillin/Streptomycin solution (G255, abm).

Incubation:

Article Title: Novel Photo- and Thermo-Responsive Nanocomposite Hydrogels Based on Functionalized rGO and Modified SIS/Chitosan Polymers for Localized Treatment of Malignant Cutaneous Melanoma
Article Snippet: .. In parallel, 100 μl of A-375 cells (American Type Culture Collection, Manassas, VA, United States) were seeded in a 96-well culture plate at a density of 1 × 106 cells/mL and incubated at 37°C, 5% CO2 for 24 h. Culture media was then removed and replaced with FBS-free DMEM. .. For each of the dilutions, 100 μl of the hydrogel extracts were added in triplicate to cell-seeded wells, and the culture plate was incubated at 37°C, 5% CO2 for another 48 h. FBS-free DMEM and DMSO (Sigma-Aldrich, St. Louis, MO, United States) were added instead of the extracts as negative and positive controls, respectively.

Derivative Assay:

Article Title: <i>In vitro</i> study of spatially-fractionated radiotherapy (SFRT) with gold nanoparticles in cancerous and healthy skin cells
Article Snippet: Purpose: This study explores how the combination of spatially-fractionated radiation therapy (SFRT) and gold nanoparticles (AuNPs) differentially affects DNA damage in melanoma and fibroblasts.. Methods and Materials: Melanoma and fibroblast monocultures and a preliminary co-culture were grown.. For AuNP conditions, cells were incubated with 10 μg/mL AuNPs.

Image Search Results

Role of four prognostically relevant genes. (A) Quantitative reverse transcription PCR for the calculated levels of four prognostically relevant genes in human epidermal melanocytes. (B) Validation of the knockdown efficiency of C2CD4B. (C, D) Scratch and Transwell assays exploring the migration and invasion of A375 cells in vitro. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, p > 0.05.

Journal: Human Mutation

Article Title: A Macrophage/Monocyte‐Related Four‐Gene Signature for Prognostic Assessment of Uveal Melanoma: BTBD6 , C2CD4B , CCL24 , and S100A4

doi: 10.1155/humu/4978880

Figure Lengend Snippet: Role of four prognostically relevant genes. (A) Quantitative reverse transcription PCR for the calculated levels of four prognostically relevant genes in human epidermal melanocytes. (B) Validation of the knockdown efficiency of C2CD4B. (C, D) Scratch and Transwell assays exploring the migration and invasion of A375 cells in vitro. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, p > 0.05.

Article Snippet: Melanoma cell line A375 (CRL‐1619) and human epidermal melanocytes (PCS‐200‐013) were purchased from American Type Culture Collection (ATCC, Manassas, Virginia) and cultured as follows.

Techniques: Reverse Transcription, Biomarker Discovery, Knockdown, Migration, In Vitro

Transcriptomic profiling of melanoma SCs identifies stemness-associated and cell line-specific molecular programs. (A) PCA of RNA-seq data from A375 and WM115 cells cultured under adherent conditions (2D) or as melanospheres (3D). Each dot represents one sample. (B) ssGSEA enrichment scores for the Melanoma stemness, WNT/β-catenin signaling, Hedgehog signaling, and Notch signaling gene sets in A375 and WM115 cells cultured in 2D or 3D conditions. Data are presented as the mean ± SEM. Each dot represents one sample. *p<0.05 by Welch’s t-test, followed by Benjamini-Hochberg correction for multiple testing across signatures. (C) Left, number of up-regulated and down-regulated DEGs identified in melanospheres compared to matched adherent cultures in A375 and WM115 cells (padj < 0.05). Right, filtered volcano plots showing DEGs (padj < 0.05, |log2FC| > 1, and baseMean > 50) in A375 and WM115 melanospheres relative to 2D cultures. The 10 most upregulated and 10 most downregulated genes are labelled. (D) Bubble plots of the top 10 most statistically significant (padj < 0.05) Reactome pathways enriched among genes up-regulated in A375 and WM115 melanospheres relative to matched 2D cultures determined by GSEA analysis. The X-axis represents the Gene Ratio and Y-axis indicates enriched pathway terms. Bubble area represents the gene count and bubble color indicates the adjusted p value. (E) VIPER analysis showing the top 15 inferred transcription factor activity in melanospheres relative to matched 2D cultures in A375 and WM115 cells. Red bars indicate transcription factors inferred to be activated in 3D cultures, whereas blue bars indicate transcription factors inferred to be activated in 2D cultures.

Journal: Frontiers in Immunology

Article Title: Melanoma stem cells drive macrophage reprogramming to a hybrid phenotype, modulating melanoma stemness and compromising NK cell-mediated immunity

doi: 10.3389/fimmu.2026.1698412

Figure Lengend Snippet: Transcriptomic profiling of melanoma SCs identifies stemness-associated and cell line-specific molecular programs. (A) PCA of RNA-seq data from A375 and WM115 cells cultured under adherent conditions (2D) or as melanospheres (3D). Each dot represents one sample. (B) ssGSEA enrichment scores for the Melanoma stemness, WNT/β-catenin signaling, Hedgehog signaling, and Notch signaling gene sets in A375 and WM115 cells cultured in 2D or 3D conditions. Data are presented as the mean ± SEM. Each dot represents one sample. *p<0.05 by Welch’s t-test, followed by Benjamini-Hochberg correction for multiple testing across signatures. (C) Left, number of up-regulated and down-regulated DEGs identified in melanospheres compared to matched adherent cultures in A375 and WM115 cells (padj < 0.05). Right, filtered volcano plots showing DEGs (padj < 0.05, |log2FC| > 1, and baseMean > 50) in A375 and WM115 melanospheres relative to 2D cultures. The 10 most upregulated and 10 most downregulated genes are labelled. (D) Bubble plots of the top 10 most statistically significant (padj < 0.05) Reactome pathways enriched among genes up-regulated in A375 and WM115 melanospheres relative to matched 2D cultures determined by GSEA analysis. The X-axis represents the Gene Ratio and Y-axis indicates enriched pathway terms. Bubble area represents the gene count and bubble color indicates the adjusted p value. (E) VIPER analysis showing the top 15 inferred transcription factor activity in melanospheres relative to matched 2D cultures in A375 and WM115 cells. Red bars indicate transcription factors inferred to be activated in 3D cultures, whereas blue bars indicate transcription factors inferred to be activated in 2D cultures.

Article Snippet: A375 human melanoma cells (CRL-1619TM), THP-1 human acute monocytic leukemia cells (TIB-202TM), NK-92 human natural killer cell line (CRL-2407TM), and K562 human chronic myelogenous leukemia cells (CCL-243TM) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: RNA Sequencing, Cell Culture, Activity Assay

Distinct secretory profiles and monocyte chemoattractant capacity of A375 and WM115 SCs. (A) PCA of multiplex ELISA assay data obtained from CM of A375 and WM115 melanospheres. Each dot represents one sample. (B) Multiplex ELISA assay quantification of soluble factors in CM from A375 and WM115 SCs. Data are presented as the mean ± SEM. Each dot represents one sample. (C) Transwell migration assay of THP-1 cells exposed to control medium, A375 SC-CM, or WM115 SC-CM, in the absence or presence of the CCR2 antagonist RS504393. Data are presented as mean ± SEM. Each dot represents one sample. Statistical analysis was performed by two-way ANOVA followed by Bonferroni-adjusted post hoc comparisons. Asterisks indicate comparisons between inhibitor-treated and untreated conditions within each medium. **p < 0.01, ***p < 0.001.

Journal: Frontiers in Immunology

Article Title: Melanoma stem cells drive macrophage reprogramming to a hybrid phenotype, modulating melanoma stemness and compromising NK cell-mediated immunity

doi: 10.3389/fimmu.2026.1698412

Figure Lengend Snippet: Distinct secretory profiles and monocyte chemoattractant capacity of A375 and WM115 SCs. (A) PCA of multiplex ELISA assay data obtained from CM of A375 and WM115 melanospheres. Each dot represents one sample. (B) Multiplex ELISA assay quantification of soluble factors in CM from A375 and WM115 SCs. Data are presented as the mean ± SEM. Each dot represents one sample. (C) Transwell migration assay of THP-1 cells exposed to control medium, A375 SC-CM, or WM115 SC-CM, in the absence or presence of the CCR2 antagonist RS504393. Data are presented as mean ± SEM. Each dot represents one sample. Statistical analysis was performed by two-way ANOVA followed by Bonferroni-adjusted post hoc comparisons. Asterisks indicate comparisons between inhibitor-treated and untreated conditions within each medium. **p < 0.01, ***p < 0.001.

Techniques: Multiplex Assay, Enzyme-linked Immunosorbent Assay, Transwell Migration Assay, Control

Transcriptomic characterization of THP-1-derived macrophages exposed to melanoma SC-CM. (A) PCA of RNA-seq data from M0 macrophages and macrophages exposed to CM from A375 or WM115 melanospheres. (B) Number of upregulated and downregulated DEGs (padj<0.05) in A375 SC-CM-educated and WM115 SC-CM-educated macrophages compared with M0 macrophages. (C) Venn diagrams showing the overlap of upregulated and downregulated DEGs between A375 SC-CM-educated and WM115 SC-CM-educated macrophages, each compared with M0 macrophages. (D) Filtered volcano plots showing DEGs (padj < 0.05, |log2FC| > 1, and baseMean > 50) in A375 SC-CM-educated versus M0 macrophages and WM115 SC-CM-educated versus M0 macrophages. The 10 most upregulated and 10 most downregulated genes are labelled. (E) Bubble plots of the top 10 most statistically significant (padj < 0.05) Reactome pathways enriched among genes upregulated in A375 SC-CM-educated macrophages and WM115 SC-CM-educated macrophages determined by GSEA analysis. The X-axis represents the Gene Ratio and Y-axis indicates enriched pathway terms. Bubble area represents the gene count, and bubble color indicates the adjusted p value. (F) VIPER analysis showing the top 15 inferred transcription factor activity in A375 SC-educated and WM115 SC-educated macrophages compared to M0 macrophages. Red bars indicate transcription factors inferred to be activated in melanoma SC-educated macrophages, whereas blue bars indicate transcription factors inferred to be activated in M0 macrophages.

Journal: Frontiers in Immunology

Article Title: Melanoma stem cells drive macrophage reprogramming to a hybrid phenotype, modulating melanoma stemness and compromising NK cell-mediated immunity

doi: 10.3389/fimmu.2026.1698412

Figure Lengend Snippet: Transcriptomic characterization of THP-1-derived macrophages exposed to melanoma SC-CM. (A) PCA of RNA-seq data from M0 macrophages and macrophages exposed to CM from A375 or WM115 melanospheres. (B) Number of upregulated and downregulated DEGs (padj<0.05) in A375 SC-CM-educated and WM115 SC-CM-educated macrophages compared with M0 macrophages. (C) Venn diagrams showing the overlap of upregulated and downregulated DEGs between A375 SC-CM-educated and WM115 SC-CM-educated macrophages, each compared with M0 macrophages. (D) Filtered volcano plots showing DEGs (padj < 0.05, |log2FC| > 1, and baseMean > 50) in A375 SC-CM-educated versus M0 macrophages and WM115 SC-CM-educated versus M0 macrophages. The 10 most upregulated and 10 most downregulated genes are labelled. (E) Bubble plots of the top 10 most statistically significant (padj < 0.05) Reactome pathways enriched among genes upregulated in A375 SC-CM-educated macrophages and WM115 SC-CM-educated macrophages determined by GSEA analysis. The X-axis represents the Gene Ratio and Y-axis indicates enriched pathway terms. Bubble area represents the gene count, and bubble color indicates the adjusted p value. (F) VIPER analysis showing the top 15 inferred transcription factor activity in A375 SC-educated and WM115 SC-educated macrophages compared to M0 macrophages. Red bars indicate transcription factors inferred to be activated in melanoma SC-educated macrophages, whereas blue bars indicate transcription factors inferred to be activated in M0 macrophages.

Techniques: Derivative Assay, RNA Sequencing, Activity Assay

ssGSEA-based inference of macrophage phenotypes induced by melanoma SC-CM. (A) Bar plots showing normalized ssGSEA scores for literature-derived TAM signatures and curated macrophage-state signatures in M0 macrophages and in macrophages exposed to A375 SC-CM or WM115 SC-CM. Curated signatures were derived from GSE115978 . Data are presented as the mean ± SEM. Each dot represents one sample. **p<0.01, ***p<0.001 by one-way ANOVA with Benjamini-Hochberg correction across signatures followed by Tukey’s multiple-comparison test. (B) Integrated matrix showing pairwise relationships among the signatures significantly modulated in vitro . The lower triangle reports Spearman correlation coefficients calculated from ssGSEA scores across samples, with asterisks indicating Benjamini-Hochberg-adjusted significance levels (*p<0.05, **p<0.01, ***p<0.001). The upper triangle reports overlap coefficients based on gene-set composition. Abbreviations: IFN, interferon-primed TAMs; Inflamm, inflammatory cytokine-enriched TAMs; Angio, pro-angiogenic TAMs; Reg, immune regulatory TAMs. (C) Bar plots representing MPI (macrophage polarization index) and AMDI (activation-induced macrophage differentiation index) values from MacSpectrum analysis of M0 macrophages and macrophages exposed to CM from A375 or WM115 melanospheres. Data are presented as the mean ± SEM. Each dot represents one sample. *p<0.05, **p<0.01 by Kruskal-Wallis test with Holm correction across the two endpoints, followed by Dunn’s post hoc test with Holm correction.

Journal: Frontiers in Immunology

Article Title: Melanoma stem cells drive macrophage reprogramming to a hybrid phenotype, modulating melanoma stemness and compromising NK cell-mediated immunity

doi: 10.3389/fimmu.2026.1698412

Figure Lengend Snippet: ssGSEA-based inference of macrophage phenotypes induced by melanoma SC-CM. (A) Bar plots showing normalized ssGSEA scores for literature-derived TAM signatures and curated macrophage-state signatures in M0 macrophages and in macrophages exposed to A375 SC-CM or WM115 SC-CM. Curated signatures were derived from GSE115978 . Data are presented as the mean ± SEM. Each dot represents one sample. **p<0.01, ***p<0.001 by one-way ANOVA with Benjamini-Hochberg correction across signatures followed by Tukey’s multiple-comparison test. (B) Integrated matrix showing pairwise relationships among the signatures significantly modulated in vitro . The lower triangle reports Spearman correlation coefficients calculated from ssGSEA scores across samples, with asterisks indicating Benjamini-Hochberg-adjusted significance levels (*p<0.05, **p<0.01, ***p<0.001). The upper triangle reports overlap coefficients based on gene-set composition. Abbreviations: IFN, interferon-primed TAMs; Inflamm, inflammatory cytokine-enriched TAMs; Angio, pro-angiogenic TAMs; Reg, immune regulatory TAMs. (C) Bar plots representing MPI (macrophage polarization index) and AMDI (activation-induced macrophage differentiation index) values from MacSpectrum analysis of M0 macrophages and macrophages exposed to CM from A375 or WM115 melanospheres. Data are presented as the mean ± SEM. Each dot represents one sample. *p<0.05, **p<0.01 by Kruskal-Wallis test with Holm correction across the two endpoints, followed by Dunn’s post hoc test with Holm correction.

Techniques: Derivative Assay, Comparison, In Vitro, Activation Assay

Functional characterization of THP-1-derived macrophages exposed to melanoma SC-CM. (A) PCA performed on multiplex ELISA assay of soluble mediator profiles from M0 macrophages and macrophages exposed to CM from A375 or WM115 melanospheres. Each dot represents one sample. (B) Multiplex ELISA assay quantification of soluble mediators secreted by M0 macrophages and by macrophages previously exposed to CM from A375 or WM115 melanospheres. Data are presented as the mean ± SEM. Each dot represents one sample. *p<0.05 by Kruskal-Wallis test with Benjamini-Hochberg correction across analytes, followed by Dunn’s post hoc test with Holm correction; pairwise p values were additionally Benjamini-Hochberg-corrected across analytes within each contrast. (C) Percentage of lysis of NK-92 cells against K562 target cells after pre-exposure to CM from M0 macrophages or from macrophages educated with A375 or WM115 SC-CM, assessed by Calcein-AM release cytotoxicity assay at the indicated E:T ratios. Data are presented as the mean ± SEM. Each dot represents one sample. The results are representative of one of at least three independent experiments showing similar results. **p<0.01, ***p<0.001 by Tukey-adjusted comparisons among experimental groups within each E:T ratio following two-way ANOVA. (D) Percentage of lysis of NK-92 cells against K562 target cells after pre-exposure to untreated or heat-treated CM from M0 macrophages or from macrophages educated with A375 or WM115 melanoma SC-CM assessed by Calcein-AM release cytotoxicity assay. Data are presented as the mean ± SEM. Each dot represents one sample. The results are representative of one of at least three independent experiments showing similar results. ***p<0.001 by Tukey-adjusted comparisons between treatment conditions within each experimental group following two-way ANOVA. (E) RT-qPCR analysis of KLF4, SOX10, POU5F1, SOX9, SOX2 in A375 and WM115 SC-enriched cultures after exposure to CM from M0 macrophages or from macrophages previously educated with melanoma SC-CM. Bar plots show mean ± SEM of 2 −ΔCt RT-qPCR values. The results are representative of one of at least three independent experiments showing similar results. *p<0.05, **p<0.01, ***p<0.001 by Welch’s t-test, followed by Holm correction for multiple testing across genes within each cell line.

Journal: Frontiers in Immunology

Article Title: Melanoma stem cells drive macrophage reprogramming to a hybrid phenotype, modulating melanoma stemness and compromising NK cell-mediated immunity

doi: 10.3389/fimmu.2026.1698412

Figure Lengend Snippet: Functional characterization of THP-1-derived macrophages exposed to melanoma SC-CM. (A) PCA performed on multiplex ELISA assay of soluble mediator profiles from M0 macrophages and macrophages exposed to CM from A375 or WM115 melanospheres. Each dot represents one sample. (B) Multiplex ELISA assay quantification of soluble mediators secreted by M0 macrophages and by macrophages previously exposed to CM from A375 or WM115 melanospheres. Data are presented as the mean ± SEM. Each dot represents one sample. *p<0.05 by Kruskal-Wallis test with Benjamini-Hochberg correction across analytes, followed by Dunn’s post hoc test with Holm correction; pairwise p values were additionally Benjamini-Hochberg-corrected across analytes within each contrast. (C) Percentage of lysis of NK-92 cells against K562 target cells after pre-exposure to CM from M0 macrophages or from macrophages educated with A375 or WM115 SC-CM, assessed by Calcein-AM release cytotoxicity assay at the indicated E:T ratios. Data are presented as the mean ± SEM. Each dot represents one sample. The results are representative of one of at least three independent experiments showing similar results. **p<0.01, ***p<0.001 by Tukey-adjusted comparisons among experimental groups within each E:T ratio following two-way ANOVA. (D) Percentage of lysis of NK-92 cells against K562 target cells after pre-exposure to untreated or heat-treated CM from M0 macrophages or from macrophages educated with A375 or WM115 melanoma SC-CM assessed by Calcein-AM release cytotoxicity assay. Data are presented as the mean ± SEM. Each dot represents one sample. The results are representative of one of at least three independent experiments showing similar results. ***p<0.001 by Tukey-adjusted comparisons between treatment conditions within each experimental group following two-way ANOVA. (E) RT-qPCR analysis of KLF4, SOX10, POU5F1, SOX9, SOX2 in A375 and WM115 SC-enriched cultures after exposure to CM from M0 macrophages or from macrophages previously educated with melanoma SC-CM. Bar plots show mean ± SEM of 2 −ΔCt RT-qPCR values. The results are representative of one of at least three independent experiments showing similar results. *p<0.05, **p<0.01, ***p<0.001 by Welch’s t-test, followed by Holm correction for multiple testing across genes within each cell line.

Techniques: Functional Assay, Derivative Assay, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Lysis, Cytotoxicity Assay, Quantitative RT-PCR

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