mouse melanoma cell lines b16bl6  (BioResource International Inc)

Journal: Biomolecules

Article Title: A Cell-Based Evaluation of the Tyrosinase-Mediated Metabolic Activation of Leukoderma-Inducing Phenols, II: The Depletion of Nrf2 Augments the Cytotoxic Effect Evoked by Tyrosinase in Melanogenic Cells

doi: 10.3390/biom15010114

Figure Lengend Snippet: Nrf2 knockdown enhances the reduction in viability of melanoma cells following their exposure to leukoderma-inducing phenolic compounds in a tyrosinase-dependent manner. B16BL6 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Nrf2 (#1, MSS275988) or Tyr (#1, MSS212191) for 24 h. ( A , D ) Levels of Nrf2 and Tyr mRNA following 24 h incubation in control medium. ( B , C , E , F ) The viability of cells following exposure to the indicated concentration of compounds for 24 h or 48 h. Data are presented as the means ± SD of independent experiments (n = 3 for ( A ) or as the number indicated for ( C , F )) or the means ± SD (n = 3 wells) with similar results in more than two independent experiments (for ( B , D , E )). *, p < 0.05, versus control siRNA-transfected cells (for ( A , D )) or between respective values. B16BL6 melanoma cells (for G – J ) were transfected with a siRNA directed against Nrf2 (#2, MSS207018), Tyr (#2, MSS212190), or a negative control siRNA ( Ctrl ) for 24 h. B16-4A5 melanoma cells (for ( K – M )) were transfected with Nrf2 (#1) and Tyr (#1). ( G , K , M ). Levels of Nrf2 or Tyr mRNA following 24 h incubation in control medium. ( H , I , J , L ) Viability of the cells following exposure to the indicated concentration of compounds for 24 h or 48 h. Data represent means ± SD (n = 3 wells). Similar results were obtained in two independent experiments (for ( H , I , J , L )). For M, data represent the means ± SD of independent experiments (as the number indicated). *, p < 0.05, between respective values or versus control siRNA-transfected cells (for ( G , K )).

Article Snippet: Mouse melanoma cell lines B16BL6 and B16 melanoma 4A5 (B16-4A5) were purchased from the RIKEN BioResource Research Center (Tsukuba, Japan) and maintained in RPMI1640 medium and Dulbecco’s modified Eagle’s medium (DMEM), respectively, containing 10% FBS.

Techniques: Knockdown, Transfection, Negative Control, Incubation, Control, Concentration Assay

Journal: Biomolecules

Article Title: A Cell-Based Evaluation of the Tyrosinase-Mediated Metabolic Activation of Leukoderma-Inducing Phenols, II: The Depletion of Nrf2 Augments the Cytotoxic Effect Evoked by Tyrosinase in Melanogenic Cells

doi: 10.3390/biom15010114

Figure Lengend Snippet: Nrf2 knockdown reduces GSH , Slc7a11, and Nqo1 mRNA levels and effectively prevents their induction by phenolic compounds. B16BL6 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Nrf2 (#1) for 24 h. The cells were further incubated in the medium containing a vehicle or compounds (0.3 mM) for 24 h. ( A , B ) Cellular GSH levels. Data represent means ± SD (n = 3 wells). Similar results were obtained in two independent experiments. ( C , D ) Levels of Slc7a11 , Nqo1 , and Nrf2 mRNA. Data represent means ± SD of independent experiments (n = 3 for ( C ), or as the number indicated for ( D )). *, p < 0.05, versus control siRNA-transfected cells; #, p < 0.05, versus vehicle-treated cells.

Article Snippet: Mouse melanoma cell lines B16BL6 and B16 melanoma 4A5 (B16-4A5) were purchased from the RIKEN BioResource Research Center (Tsukuba, Japan) and maintained in RPMI1640 medium and Dulbecco’s modified Eagle’s medium (DMEM), respectively, containing 10% FBS.

Techniques: Knockdown, Transfection, Negative Control, Incubation, Control

Journal: Biomolecules

Article Title: A Cell-Based Evaluation of the Tyrosinase-Mediated Metabolic Activation of Leukoderma-Inducing Phenols, II: The Depletion of Nrf2 Augments the Cytotoxic Effect Evoked by Tyrosinase in Melanogenic Cells

doi: 10.3390/biom15010114

Figure Lengend Snippet: Slc7a11 knockdown enhances the cytotoxicity induced by RD, RK, and pCRE, whereas Nqo1 knockdown or inhibition is ineffective for most compounds. B16BL6 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Slc7a11 or Nqo1 for 24 h. ( A ) Slc7a11 and Nqo1 mRNA levels following 24 h incubation in a control medium. ( B – D ) The viability of cells following exposure to the indicated concentration of compounds for 24 h or 48 h. ( E , F ) The viability of B16BL6 cells after exposure to the indicated concentration of phenolic compounds in the presence or absence of ES936 (500 nM) for 24 h. ( F ) Cells were pretreated with a vehicle or ES936 for 24 h before their exposure to the compounds with/without ES936. ( G ) Western blots and quantitation of tyrosinase and the loading control of β-actin in cells at 48 h after Nqo1 -siRNA transfection or 24 h after ES936 treatment. Data represent means ± SD (n = 3 wells) with similar results in two independent experiments (for ( A – F )) or means ± SD of independent experiments (n = 4 for RD in panel C, D or n = 3 for 4SCAP in panel D). *, p < 0.05, versus Ctrl -siRNA-transfected cells (for ( A )) or between respective values (for ( B – F )). B16-4A5 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Slc7a11 or Nqo1 for 24 h. ( H ) Levels of Slc7a11 and Nqo1 mRNA following 24 h incubation in a control medium. ( I ) The viability of cells following exposure to the indicated concentration of compounds for 24 h or 48 h. Data represent means ± SD (n = 3 wells). Similar results were obtained in two independent experiments (for ( I )). *, p < 0.05, versus Ctrl -siRNA-transfected cells.

Article Snippet: Mouse melanoma cell lines B16BL6 and B16 melanoma 4A5 (B16-4A5) were purchased from the RIKEN BioResource Research Center (Tsukuba, Japan) and maintained in RPMI1640 medium and Dulbecco’s modified Eagle’s medium (DMEM), respectively, containing 10% FBS.

Techniques: Knockdown, Inhibition, Transfection, Negative Control, Incubation, Control, Concentration Assay, Western Blot, Quantitation Assay