Journal: Biomedicines

Article Title: Apoptosis, Cell Growth, and Glycogen Synthase Kinase 3β Phosphorylation in Caffeic Acid-Treated Human Malignant Melanoma Cells

doi: 10.3390/biomedicines13102389

Figure Lengend Snippet: Cytotoxic effects of CA on G361 cells. ( A ) Chemical structure of caffeic acid. ( B ) G361 and SK-MEL-24 cells were treated with various concentrations of CA (0, 80, 160, 240, and 320 μM) for 24, 48, and 72 h. Cell viability was measured using the MTT assay. IC 50 values for G361 cells were calculated for each time point. * p < 0.05 versus control. ( C ) Intracellular ATP levels were measured after treatment with CA (0, 20, 40, and 80 μM) for 24 h. ( D ) Phase contrast microscopy images of G361 cells treated with CA (0, 20, 40, and 80 μM) for 24 h. Scale bar: 50 μm. ( E ) Nuclear morphology was assessed using DAPI staining after 24 h of CA treatment. Apoptotic nuclear condensation and fragmentation were observed in a dose-dependent manner. Scale bar: 25 μm. ( F ) Quantification of apoptotic cells using the Annexin V-PE binding assay and Muse Cell Analyzer after 24 h of CA treatment.

Article Snippet: The human malignant melanoma cell line G361 and SK-MEL-24 was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: MTT Assay, Control, Microscopy, Staining, Binding Assay

Journal: Development & Reproduction

Article Title: Antitumor Effect of Metformin in Combination with Binimetinib on Melanoma Cells

doi: 10.12717/DR.2021.25.2.93

Figure Lengend Snippet: (A) Dose-response curves for metformin (Metf), binimetinib (Bini) and their combination (B+M) in constant ratio (40:1) (n=3, error bar represents SE). G361 cells were seeded at 5×10 4 cells/well (0.5 mL) in 24-well culture plates, incubated for 24 h and then treated with metformin, binimetinib and their combination for 72 h. MTT assay was performed for the determination of cell viability. The viability of control cells was regarded as 100%. (B) Combination index (CI) values for the various combination points of metformin and trametinib. The CI values were calculated by using CompuSyn software. (C) Western blot analysis for the activity of ERK. Cells were treated with increasing concentrations of metformin and binimetinib for 24 h. β-actin was used as a loading control.

Article Snippet: We purchased the human melanoma cell line G361 from Korean Cell Line Bank (Seoul, Korea).

Techniques: Incubation, MTT Assay, Control, Software, Western Blot, Activity Assay

Journal: The Journal of Clinical Investigation

Article Title: H3K27me3-mediated PGC1 α gene silencing promotes melanoma invasion through WNT5A and YAP

doi: 10.1172/JCI130038

Figure Lengend Snippet: (A and B) Overexpression of PGC1α by adenovirus suppresses YAP protein expression and activity without affecting its mRNA transcript in A375P, A375 (A), and G361 (B) melanoma cells (n = 3). (C and D) Depletion of PGC1α by shRNA (C) or CRISPR (D) enhances YAP protein expression and activation without altering its mRNA level in multiple melanoma cell lines (n = 3). (E) YAP protein is more stable in A375P cells with PGC1α depletion. Cells were treated with 50 mg/mL of CHX for the indicated period and harvested for immunoblotting analysis (n = 3). (F) PGC1α-induced YAP degradation in A375P cells can be blocked by treatment with 10 μM of proteasome inhibitor MG132 overnight (n = 3). Quantitative results were analyzed by Student’s t test or 1-way ANOVA. Data are shown as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.005.

Article Snippet: A total of 2 × 10 6 A375P cells or 1 × 10 6 G361 cells in 0.2 mL of DMEM were injected into the tail veins of 8-week-old NCI Ath/Nu mice (Charles River).

Techniques: Over Expression, Expressing, Activity Assay, shRNA, CRISPR, Activation Assay, Western Blot

Journal: The Journal of Clinical Investigation

Article Title: H3K27me3-mediated PGC1 α gene silencing promotes melanoma invasion through WNT5A and YAP

doi: 10.1172/JCI130038

Figure Lengend Snippet: (A) In highly invasive, PGC1α-negative A375 melanoma cells, depletion of YAP by shRNA compromises its migratory ability (n = 3). (B) In the highly migratory murine melanoma cell line B16BL6, depletion of YAP by shRNA suppresses its invasiveness in congenic mice (n = 3 for qPCR and n = 5 mice per group). (C and D) In PGC1α-positive A375P and G361 melanoma cells, the increased metastasis by loss of PGC1α can be prevented by depletion of YAP, as measured by in vivo lung colonization assay (n = 6–16 mice per group). Quantitative results were analyzed by Student’s t test (A and B) or 1-way ANOVA (D). Data are shown as mean ± SEM if not otherwise indicated. *P < 0.05; **P < 0.01; ***P < 0.005.

Article Snippet: A total of 2 × 10 6 A375P cells or 1 × 10 6 G361 cells in 0.2 mL of DMEM were injected into the tail veins of 8-week-old NCI Ath/Nu mice (Charles River).

Techniques: shRNA, In Vivo