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human lymph node metastatic melanoma cell line sk mel 28 (ATCC)

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ATCC human lymph node metastatic melanoma cell line sk mel 28
Human Lymph Node Metastatic Melanoma Cell Line Sk Mel 28, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 2204 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/metastatic+melanoma/SK-MEL-28/10__3390_slash_antiox15060740-69-11-19
Average 99 stars, based on 2204 article reviews

human lymph node metastatic melanoma cell line sk mel 28 - by Bioz Stars, 2026-09

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Multiple Displacement Amplification:

Article Title: MFF Regulation of Mitochondrial Cell Death Is a Therapeutic Target in Cancer
Article Snippet: .. Human prostate adenocarcinoma (LNCaP, C4-2, C4-2B, PC3, DU145), NSCLC (A549, H460), breast adenocarcinoma (MDA-231, MCF7), metastatic melanoma (A2058), normal prostate epithelial (RWPE-1) and human glioblastoma (LN229) cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA), and maintained in culture according to the supplier’s specifications. ..

Article Title: MFF REGULATION OF MITOCHONDRIAL CELL DEATH IS A THERAPEUTIC TARGET IN CANCER
Article Snippet: .. Cells and cell culture Human prostate adenocarcinoma (LNCaP, C4–2, C4–2B, PC3, DU145), NSCLC (A549, H460), breast adenocarcinoma (MDA-231, MCF7), metastatic melanoma (A2058), normal prostate epithelial (RWPE-1) and human glioblastoma (LN229) cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA), and maintained in culture according to the supplier’s specifications. ..

Article Title: MFF REGULATION OF MITOCHONDRIAL CELL DEATH IS A THERAPEUTIC TARGET IN CANCER
Article Snippet: .. Human prostate adenocarcinoma (LNCaP, C4–2, C4–2B, PC3, DU145), NSCLC (A549, H460), breast adenocarcinoma (MDA-231, MCF7), metastatic melanoma (A2058), normal prostate epithelial (RWPE-1) and human glioblastoma (LN229) cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA), and maintained in culture according to the supplier’s specifications. ..

Cell Culture:

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Efficiency of lentiviral transduction with il12, il15 and il15Rα or il18 genes and the level of differentiation of bone marrow-derived dendritic cells (DCs) stimulated with B16 F0 tumor antigens (TAg). Concentration of overexpressed cytokines IL-12 (A) , IL-15 (B) , IL-18 (C) in supernatants collected after 48 hours of DCs cultures measured using ELISA. Percentage of CD11c + cells on the 10 th day of DCs cultured (D) . Expression of CD40 (E) , CD80 (F) , CD86 (G) and MHC II (H) molecules on the surface of CD11c + cells (MFI) (bar plots and representative overlay histograms). Results are presented as mean+SD calculated for 5–6 samples per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (A–D, F–H) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (E) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the DC/TAg control cells; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg control cells, asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ x p<0.05; **/ xx p<0.01; ***/ xxx p<0.001; ****/ xxxx p<0.0001). IC – isotype control , MFI – mean fluorescence intensity .

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: Efficiency of lentiviral transduction with il12, il15 and il15Rα or il18 genes and the level of differentiation of bone marrow-derived dendritic cells (DCs) stimulated with B16 F0 tumor antigens (TAg). Concentration of overexpressed cytokines IL-12 (A) , IL-15 (B) , IL-18 (C) in supernatants collected after 48 hours of DCs cultures measured using ELISA. Percentage of CD11c + cells on the 10 th day of DCs cultured (D) . Expression of CD40 (E) , CD80 (F) , CD86 (G) and MHC II (H) molecules on the surface of CD11c + cells (MFI) (bar plots and representative overlay histograms). Results are presented as mean+SD calculated for 5–6 samples per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (A–D, F–H) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (E) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the DC/TAg control cells; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg control cells, asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ x p<0.05; **/ xx p<0.01; ***/ xxx p<0.001; ****/ xxxx p<0.0001). IC – isotype control , MFI – mean fluorescence intensity .

Article Snippet: The non-metastatic murine melanoma B16 F0 cell line (ECACC 92101204) was cultured in high glucose Dulbecco’s Modified Eagle Medium (DMEM, ATCC) supplemented with 100 U/ml penicillin, 100 mg/ml streptomycin and 10% heat-inactivated fetal bovine serum (FBS; Sigma−Aldrich).

Techniques: Transduction, Derivative Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Control, Fluorescence

The ability of DCs genetically modified to produce IL-12, IL-15/Il-15Rα or IL-18 and stimulated with B16 F0 tumor antigens to prime splenocytes. Percentage of CD8 + cells (A) , CD4 + cells (B) and NK cells (C) among splenocytes obtained after 5-day coculture with DCs. Percentage of effector cells (CD107a + ) among CD8 + (D) , CD4 + (E) and NK cells (F) after 2-hour incubation with B16 F0 cells. Concentration of IFN-γ (G) and IL-10 (H) in supernatants collected after 5 days of cocultured DCs and spleen cells. Ratio of IFN-γ and IL-10 concentration (I) . Cytotoxic activity of splenocytes presented as a percentage of dead B16 F0 tumor cells after 4-hour incubation with effector cells in a ratio of 10:1 (effector:target) (J) . Results are presented as mean+SD calculated for 6–12 samples per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (A–D, G–I) , the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (E, J) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (F) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the DC/TAg control cells; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg control cells, asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ x p<0.05; **/ xx p<0.01; ***/ xxx p<0.001; ****/ xxxx p<0.0001).

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: The ability of DCs genetically modified to produce IL-12, IL-15/Il-15Rα or IL-18 and stimulated with B16 F0 tumor antigens to prime splenocytes. Percentage of CD8 + cells (A) , CD4 + cells (B) and NK cells (C) among splenocytes obtained after 5-day coculture with DCs. Percentage of effector cells (CD107a + ) among CD8 + (D) , CD4 + (E) and NK cells (F) after 2-hour incubation with B16 F0 cells. Concentration of IFN-γ (G) and IL-10 (H) in supernatants collected after 5 days of cocultured DCs and spleen cells. Ratio of IFN-γ and IL-10 concentration (I) . Cytotoxic activity of splenocytes presented as a percentage of dead B16 F0 tumor cells after 4-hour incubation with effector cells in a ratio of 10:1 (effector:target) (J) . Results are presented as mean+SD calculated for 6–12 samples per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (A–D, G–I) , the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (E, J) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (F) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the DC/TAg control cells; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg control cells, asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ x p<0.05; **/ xx p<0.01; ***/ xxx p<0.001; ****/ xxxx p<0.0001).

Techniques: Genetically Modified, Incubation, Concentration Assay, Activity Assay, Control

Growth of B16 F0 tumors in mice after immunotherapy and chemoimmunotherapy composed of a chemotherapeutic agent, anti-IL-10R antibody, and genetically modified DC-based vaccines stimulated with B16 F0 tumor antigens. Scheme of immunotherapy (A) and chemoimmunotherapy (E) treatment created with BioRender.com . Growth kinetics of B16 F0 tumor in mice treated with immunotherapy (B) or chemoimmunotherapy (F) . Violin plot presenting individual tumor volume and designated median tumor volume for each group, calculated on the 19 th day of the immunotherapy (C) or chemoimmunotherapy (G) experiment. Results are presented as median for 3–10 mice per group. Table presenting B16 F0 tumor growth inhibition (TGI) calculated on the 19 th day of the experiment in relation to the non-treated group (nt) (D, H) . Differences between groups were estimated using the two−way ANOVA followed by Tukey’s multiple comparisons post−hoc test (B, F) or non-parametric Kruskal−Wallis test followed by Dunn’s multiple comparisons test (C, G) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M) – (*p<0.05; **/ ## p<0.01; ***p<0.001; ****p<0.0001).

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: Growth of B16 F0 tumors in mice after immunotherapy and chemoimmunotherapy composed of a chemotherapeutic agent, anti-IL-10R antibody, and genetically modified DC-based vaccines stimulated with B16 F0 tumor antigens. Scheme of immunotherapy (A) and chemoimmunotherapy (E) treatment created with BioRender.com . Growth kinetics of B16 F0 tumor in mice treated with immunotherapy (B) or chemoimmunotherapy (F) . Violin plot presenting individual tumor volume and designated median tumor volume for each group, calculated on the 19 th day of the immunotherapy (C) or chemoimmunotherapy (G) experiment. Results are presented as median for 3–10 mice per group. Table presenting B16 F0 tumor growth inhibition (TGI) calculated on the 19 th day of the experiment in relation to the non-treated group (nt) (D, H) . Differences between groups were estimated using the two−way ANOVA followed by Tukey’s multiple comparisons post−hoc test (B, F) or non-parametric Kruskal−Wallis test followed by Dunn’s multiple comparisons test (C, G) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M) – (*p<0.05; **/ ## p<0.01; ***p<0.001; ****p<0.0001).

Techniques: Genetically Modified, Vaccines, Inhibition, Control

Evaluation of leukocyte subpopulations infiltrating B16 F0 tumor tissue after administration of immunotherapy or chemoimmunotherapy. Scheme of the multiparameter flow cytometry analysis of myeloid and lymphoid cells infiltrating B16 F0 tumor nodules (prepared for one representative sample from group IX in the chemoimmunotherapy experiment) (A) . Percentage of live CD45 + cells in tumor (B, H) . Percentage of each leukocyte’s population among CD45 + cells (C, I) . Percentage of CD8 + (D, J) , CD4 + (E, K) , Treg (F, L) and NK cells (G, M) infiltrating tumor tissue. Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (B, G, J, M) , the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (D) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (K, L) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M) ; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # / x p<0.05; **/ xx p<0.01; ***/ xxx p<0.001; ****p<0.0001).

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: Evaluation of leukocyte subpopulations infiltrating B16 F0 tumor tissue after administration of immunotherapy or chemoimmunotherapy. Scheme of the multiparameter flow cytometry analysis of myeloid and lymphoid cells infiltrating B16 F0 tumor nodules (prepared for one representative sample from group IX in the chemoimmunotherapy experiment) (A) . Percentage of live CD45 + cells in tumor (B, H) . Percentage of each leukocyte’s population among CD45 + cells (C, I) . Percentage of CD8 + (D, J) , CD4 + (E, K) , Treg (F, L) and NK cells (G, M) infiltrating tumor tissue. Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (B, G, J, M) , the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (D) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (K, L) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M) ; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # / x p<0.05; **/ xx p<0.01; ***/ xxx p<0.001; ****p<0.0001).

Techniques: Flow Cytometry, Control

Estimation of TAM and MDSC subpopulation in B16 F0 tumor tissue after applied therapy. Percentage of TAM (A, D) , TAMs MHC II high /TAMs MHC II low (B, E) and MDSC (C, F) among CD45 + cells in tumors. Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (A, D) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (F) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M); crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # p<0.05; **/ ## / xx p<0.01; ***/ xxx p<0.001; ****/ #### p<0.0001).

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: Estimation of TAM and MDSC subpopulation in B16 F0 tumor tissue after applied therapy. Percentage of TAM (A, D) , TAMs MHC II high /TAMs MHC II low (B, E) and MDSC (C, F) among CD45 + cells in tumors. Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (A, D) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (F) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M); crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # p<0.05; **/ ## / xx p<0.01; ***/ xxx p<0.001; ****/ #### p<0.0001).

Techniques: Control

Impact of applied immunotherapy and chemoimmunotherapy on the induction of systemic antitumor in B16 F0 melanoma model. Scheme of the flow cytometry analysis of restimulated splenocytes (A) . Percentage of CD8 + (B, H) , CD4 + (C, I) , and NK cells (D, J) restimulated splenocytes. Percentage of CD107a + cells among CD8 + (E, K) , CD4 + (F, L) and NK cells (G, M) . Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (E, H, K) , the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (B, F, G, I) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (C, D, J, L, M) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M) ; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # p<0.05; **/ ## / xx p<0.01; ***/ ### p<0.001; ****/ #### p<0.0001).

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: Impact of applied immunotherapy and chemoimmunotherapy on the induction of systemic antitumor in B16 F0 melanoma model. Scheme of the flow cytometry analysis of restimulated splenocytes (A) . Percentage of CD8 + (B, H) , CD4 + (C, I) , and NK cells (D, J) restimulated splenocytes. Percentage of CD107a + cells among CD8 + (E, K) , CD4 + (F, L) and NK cells (G, M) . Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (E, H, K) , the parametric one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test (B, F, G, I) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (C, D, J, L, M) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M) ; crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # p<0.05; **/ ## / xx p<0.01; ***/ ### p<0.001; ****/ #### p<0.0001).

Techniques: Flow Cytometry, Control

Activity of restimulated splenocytes obtained from B16 F0 melanoma-bearing mice after immunotherapy and chemoimmunotherapy. Concentration of IFN-γ (A, D) , IL-10 (B, E) and IL-4 (C, F) in supernatants after restimulation of spleen cells with B16 F0 cells, measured using ELISA assay. Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (A, C, F) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (B, D, E) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M); crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # p<0.05; **/ ## p<0.01; ***/ ### p<0.001; ****/ #### p<0.0001).

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: Activity of restimulated splenocytes obtained from B16 F0 melanoma-bearing mice after immunotherapy and chemoimmunotherapy. Concentration of IFN-γ (A, D) , IL-10 (B, E) and IL-4 (C, F) in supernatants after restimulation of spleen cells with B16 F0 cells, measured using ELISA assay. Results are presented as mean+SD calculated for 3–6 mice per group. Differences between groups were estimated using the non-parametric Kruskal-Wallis test followed by Dunn’s multiple comparisons post-hoc test (A, C, F) or the parametric Brown-Forsythe and Welch ANOVA test followed by Dunnett’s T3 multiple comparisons post-hoc test (B, D, E) . The asterisks (*) presented in the graphs indicate statistically significant differences between the given groups and the non-treated control group (nt); hashtags (#) above a bar indicate a statistically significant difference between the given group and the HES-MTX treated group (H-M); crosses (X) indicate a statistically significant difference between the given group and the DC/Vctrl/TAg treated group; asterisks (*) under the line indicate statistically significant differences between the given groups – (*/ # p<0.05; **/ ## p<0.01; ***/ ### p<0.001; ****/ #### p<0.0001).

Techniques: Activity Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

The effect of chemoimmunotherapy with DC/IL-12/TAg + DC/IL-15/IL-15Rα/TAg + DC/IL-18/TAg cell vaccine on the inhibition of B16 F0 tumor growth.

Journal: Frontiers in Immunology

Article Title: Co-delivery of IL-12/IL-15/IL-18 engineered DC vaccines with anti-IL-10R and nanoconjugated methotrexate in melanoma

doi: 10.3389/fimmu.2026.1773836

Figure Lengend Snippet: The effect of chemoimmunotherapy with DC/IL-12/TAg + DC/IL-15/IL-15Rα/TAg + DC/IL-18/TAg cell vaccine on the inhibition of B16 F0 tumor growth.

Techniques: Inhibition

SLC45A2 knockdown inhibits cell migration and clonogenicity in uveal melanoma C918 and MuM2B cells. (A, B) qRT-PCR and Western blotting to evaluate the level of SLC45A2 mRNA and protein levels 48 h after transfection. All siRNA sequences could result in significant decrease in SLC45A2 mRNA and protein levels expression (P<0.001). (C) Transwell assay. The migration and invasion capacity of C918 and MuM2B cells decreased significantly after SLC45A2 knockdown. (D) Scratch-wound healing assay. A significantly slower wound healing rate was observed in cells with a decreased expression of SLC45A2 gene. All data were presented as the means ± SD of three independent experiments. (*P < 0.05, **P < 0.01, ***P < 0.001).

Journal: Frontiers in Immunology

Article Title: Single-cell profiling deciphering cholesterol metabolism dysregulation in metastatic uveal melanoma and implicating SLC45A2 in its prognosis

doi: 10.3389/fimmu.2025.1660268

Figure Lengend Snippet: SLC45A2 knockdown inhibits cell migration and clonogenicity in uveal melanoma C918 and MuM2B cells. (A, B) qRT-PCR and Western blotting to evaluate the level of SLC45A2 mRNA and protein levels 48 h after transfection. All siRNA sequences could result in significant decrease in SLC45A2 mRNA and protein levels expression (P<0.001). (C) Transwell assay. The migration and invasion capacity of C918 and MuM2B cells decreased significantly after SLC45A2 knockdown. (D) Scratch-wound healing assay. A significantly slower wound healing rate was observed in cells with a decreased expression of SLC45A2 gene. All data were presented as the means ± SD of three independent experiments. (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Briefly, Human metastatic uveal melanoma cell line C918 (CL-0264 Procell Inc., Wuhan, China) and MuM2B (FH-1158 Fuheng Biotechnology Inc., Shanghai, China) was cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum.

Techniques: Knockdown, Migration, Quantitative RT-PCR, Western Blot, Transfection, Expressing, Transwell Assay, Wound Healing Assay

(A) Schematic representation of the zebrafish xenograft assay. (B) Migration capacity was assessed in MeWo-GFP or A375P-GFP cells. Starting 48h after siRNA transfection, cell migration was tracked by live microscopy for 16 h. Migration velocity was reduced in both cell lines following PFKFB4 knockdown with siRNA targeting PFKFB4 (siPFKFB4), compared with cells transfected with non-targeting control siRNA (siCtrl). Bar plots represent mean ± SEM of three independent experiments (unpaired t- test). Kernel density plots illustrate the distribution of migration speeds in siCtrl versus siPFKFB4 groups. (C) (E) Number of invasive MeWo-GFP foci (C) or A375P-GFP foci (E) per fish at 3 days post injection (dpi) after transfection with siCtrl or siPFKFB4. (D) (F) Representative zebrafish larvae at 3 dpi xenografted with MeWo-GFP (D) or A375P-GFP (F) cells after transfection with siCtrl or siPFKFB4. Asterisks indicate invasive foci detached from the primary tumor. Orange dotted lines mark the swim bladder boundary. (G) Number of distant metastatic A375P-GFP foci, defined as cells migrating > 300 μm from the tumor boundary. (H) Magnification of the tail region showing A375P-GFP cells metastasizing to caudal hematopoietic tissue. In panels C , E , and G , each dot represents one fish. Statistical significance was determined by unpaired t- test. n.s., not significant (P > 0.05); *P < 0.05; dpi, days post injection; dpf, days post fertilization.

Journal: bioRxiv

Article Title: Zebrafish Xenografts Reveal a Context-dependent Role of PFKFB4 in Melanoma Cell

doi: 10.1101/2025.09.06.674616

Figure Lengend Snippet: (A) Schematic representation of the zebrafish xenograft assay. (B) Migration capacity was assessed in MeWo-GFP or A375P-GFP cells. Starting 48h after siRNA transfection, cell migration was tracked by live microscopy for 16 h. Migration velocity was reduced in both cell lines following PFKFB4 knockdown with siRNA targeting PFKFB4 (siPFKFB4), compared with cells transfected with non-targeting control siRNA (siCtrl). Bar plots represent mean ± SEM of three independent experiments (unpaired t- test). Kernel density plots illustrate the distribution of migration speeds in siCtrl versus siPFKFB4 groups. (C) (E) Number of invasive MeWo-GFP foci (C) or A375P-GFP foci (E) per fish at 3 days post injection (dpi) after transfection with siCtrl or siPFKFB4. (D) (F) Representative zebrafish larvae at 3 dpi xenografted with MeWo-GFP (D) or A375P-GFP (F) cells after transfection with siCtrl or siPFKFB4. Asterisks indicate invasive foci detached from the primary tumor. Orange dotted lines mark the swim bladder boundary. (G) Number of distant metastatic A375P-GFP foci, defined as cells migrating > 300 μm from the tumor boundary. (H) Magnification of the tail region showing A375P-GFP cells metastasizing to caudal hematopoietic tissue. In panels C , E , and G , each dot represents one fish. Statistical significance was determined by unpaired t- test. n.s., not significant (P > 0.05); *P < 0.05; dpi, days post injection; dpf, days post fertilization.

Article Snippet: Human metastatic melanoma cell lines MeWo (ATCC #HTB-65) , A375M , and A375P-GFP (ATCC #CRL-3224), as well as HEK-293FT cells (Thermo Fisher), were maintained in complete RPMI-1640 medium (MeWo, A375M; Gibco, #21875-034) or in complete DMEM (A375P, HEK-293FT; Gibco, # 41965-039), supplemented with 10% fetal bovine serum (Eurobio, #CVFSVF00-0U) and 1% penicillin/streptomycin (Sigma, #P0781).

Techniques: Xenograft Assay, Migration, Transfection, Microscopy, Knockdown, Control, Injection

(A) Protein expression levels of the EMT-TFs SNAIL2, SNAIL1, TWIST, ZEB2 and ZEB1 across different melanoma cell lines. (B) mRNA expression of EMT-TFs in MeWo cells assessed by qPCR 48h after transfection with siCtrl or siPFKFB4. Statistical analysis was performed using an unpaired t-test. (C, D) Normalized protein expression of EMT-TFs in MeWo (C) or A375M (D) cells, 48h after transfection with siCtrl or siPFKFB4. Each dot represents one biological replicate with paired values connected by lines. Statistical analysis was performed using a paired t- test. n.s, not significant (P > 0.05); *P < 0.05; **P < 0.01.

Journal: bioRxiv

Article Title: Zebrafish Xenografts Reveal a Context-dependent Role of PFKFB4 in Melanoma Cell

doi: 10.1101/2025.09.06.674616

Figure Lengend Snippet: (A) Protein expression levels of the EMT-TFs SNAIL2, SNAIL1, TWIST, ZEB2 and ZEB1 across different melanoma cell lines. (B) mRNA expression of EMT-TFs in MeWo cells assessed by qPCR 48h after transfection with siCtrl or siPFKFB4. Statistical analysis was performed using an unpaired t-test. (C, D) Normalized protein expression of EMT-TFs in MeWo (C) or A375M (D) cells, 48h after transfection with siCtrl or siPFKFB4. Each dot represents one biological replicate with paired values connected by lines. Statistical analysis was performed using a paired t- test. n.s, not significant (P > 0.05); *P < 0.05; **P < 0.01.

Techniques: Expressing, Transfection

(A) Mean migration velocity of MeWo cells 48 h after transfection with siCtrl or siRNA targeting SNAIL2 (siSNAIL2). Kernel density plots illustrate the distribution of migration speeds in control versus SNAIL2 knockdown groups. (B,C) Expression of PFKFB4 and SNAIL2 proteins (B) and mean migration velocity (C) of MeWo cells transfected with siCtrl or siRNA targeting PFKFB4 (siPFKFB4), combined with empty pcDNA vector or a plasmid expressing Xenopus laevis SNAIL2 (xSNAIL2). Kernel density plots show the distribution of migration speeds across groups. xSNAIL2 failed to rescue the migration defect induced by PFKFB4 knockdown. (D) Mean migration velocity of MeWo cells after siCtrl or siPFKFB4 transfection, combined with empty pcDNA vector or a plasmid expressing Xenopus laevis PFKFB4 (xPFKFB4). Kernel density plots show the distribution of migration speeds. xPFKFB4 was able to rescue the migration defect caused by PFKFB4 knockdown. (E, F) Protein expression levels of PFKFB4 and SNAIL2 (E) and quantification from four biological replicates (F) in MeWo cells transfected with siCtrl or siPFKFB4, combined with empty pcDNA or xPFKFB4 plasmid. xPFKFB4 failed to rescue the decrease in SNAIL2 protein levels caused by PFKFB4 knockdown. In panels A , C , and D , bar plots represent mean ±SEM and were analyzed using an unpaired t- test. (F) Each dot represents one biological replicate, with paired values connected by lines; significance was assessed using a paired t- test. n.s., not significant (P > 0.05); *P < 0.05.

Journal: bioRxiv

Article Title: Zebrafish Xenografts Reveal a Context-dependent Role of PFKFB4 in Melanoma Cell

doi: 10.1101/2025.09.06.674616

Figure Lengend Snippet: (A) Mean migration velocity of MeWo cells 48 h after transfection with siCtrl or siRNA targeting SNAIL2 (siSNAIL2). Kernel density plots illustrate the distribution of migration speeds in control versus SNAIL2 knockdown groups. (B,C) Expression of PFKFB4 and SNAIL2 proteins (B) and mean migration velocity (C) of MeWo cells transfected with siCtrl or siRNA targeting PFKFB4 (siPFKFB4), combined with empty pcDNA vector or a plasmid expressing Xenopus laevis SNAIL2 (xSNAIL2). Kernel density plots show the distribution of migration speeds across groups. xSNAIL2 failed to rescue the migration defect induced by PFKFB4 knockdown. (D) Mean migration velocity of MeWo cells after siCtrl or siPFKFB4 transfection, combined with empty pcDNA vector or a plasmid expressing Xenopus laevis PFKFB4 (xPFKFB4). Kernel density plots show the distribution of migration speeds. xPFKFB4 was able to rescue the migration defect caused by PFKFB4 knockdown. (E, F) Protein expression levels of PFKFB4 and SNAIL2 (E) and quantification from four biological replicates (F) in MeWo cells transfected with siCtrl or siPFKFB4, combined with empty pcDNA or xPFKFB4 plasmid. xPFKFB4 failed to rescue the decrease in SNAIL2 protein levels caused by PFKFB4 knockdown. In panels A , C , and D , bar plots represent mean ±SEM and were analyzed using an unpaired t- test. (F) Each dot represents one biological replicate, with paired values connected by lines; significance was assessed using a paired t- test. n.s., not significant (P > 0.05); *P < 0.05.

Techniques: Migration, Transfection, Control, Knockdown, Expressing, Plasmid Preparation

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