Fig. 4

Roles of TPPP3, DOCK2, EIF3H, RNF128, DAPK1, and SYT7 in apoptosis of melanoma stem cells. a Expression levels of mRNAs in melanoma stem cell and non-stem cells. Northern blot was conducted to detect the mRNA level. β-tubulin was used as a control. b Detection of proteins in melanoma stem cells and non-stem cells by Western blot. β-tubulin was used as a control. c Silencing of TPPP3, DOCK2, EIF3H, RNF128, DAPK1, or SYT7 in melanoma stem cells. Melanoma stem cells were transfected with sequence-specific siRNA. At 36 h after transfection, the expression level of mRNA was detected using quantitative real-time PCR. As a control, siRNA-scrambled was included in the assays (**p < 0.01). d Western blot analysis of gene silencing in melanoma stem cells. e Influence of gene silencing on apoptosis of melanoma stem cells. Melanoma stem cells treated with siRNA were evaluated by caspase 3/7 activity assay at 36 h after siRNA transfection (*p < 0.05, **p < 0.01). f Overexpression of TPPP3, DOCK2, EIF3H, RNF128, DAPK1, or SYT7 in melanoma stem cells. Melanoma stem cells were transfected with the recombinant plasmid expressing TPPP3, DOCK2, EIF3H, RNF128, DAPK1, or SYT7. As a control, vector alone was included in the transfection. At 48 h after transfection, the proteins were examined with Western blot. g Impact of gene overexpression on apoptosis of melanoma stem cells. Melanoma stem cells transfected with a vector expressing a gene were subjected to caspase 3/7 activity assay at 48 h after transfection (**p < 0.01). Vector alone was used as a control. All the experiments were biologically repeated three times. h Mode for the hnRNP A2B1-mediated apoptosis suppression of melanoma stem cells