Fig. 6
Effect of lj-1-59 on BRAFi-resistant melanoma cells. a BRAFi-resistant melanoma cells (RA) were generated as described in “Methods”. RA (left panel) and parental A375 (right panel) cells were prepared in 96-well plates. The cells were treated with PLX4032. Cell viability was determined by CCK-8 assay. The results represent the means (n = 6) ± SD, and asterisk (*) indicates a significant difference (p < 0.05, Student’s t-test). b RA cells were treated with increasing dose lj-1-59 for 0-72 h (left panel). Cell viability was determined by CCK-8 assay. The results represent the means (n = 6) ± SD, and asterisk (*) indicates a significant difference (p < 0.05, Student’s t-test). The IC50 values of lj-1-59 in RA cells were automatically calculated by GraphPad Prism software (right panel). c RA cells were prepared in 6-well plates. The cells were treated with increasing dose lj-1-59 for 24 h. After 2 weeks, the number of colonies was assessed and quantified as described in “Methods”. The results represent the means (n = 5) ± SD, and asterisk (*) indicates a significant difference (p < 0.05, Student’s t-test). d Cell cycle analysis of RA cells with increasing dose lj-1-59 for the 48 h. The cell cycle distribution was detected by flow cytometry as described in “Methods”. The results are expressed as the means (n = 4) ± SD, and asterisk (*) indicates a significant difference (p < 0.05, Chi-square). e RA cells were treated with increasing dose lj-1-59 for the 48 h. Apoptosis was detected by flow cytometry as described in “Methods”. The results are expressed as the means (n = 4) ± SD, and asterisk (*) indicates a significant difference (p < 0.05, Student’s t-test). f Western Blot analysis of apoptosis-associated proteins in RA cells with lj-1-59 treatment for 48 h. GAPDH was used as a loading control