Fig. 3
From: Requirement of splicing factor hnRNP A2B1 for tumorigenesis of melanoma stem cells
Underlying mechanism of hnRNP A2B1 on the stemness of melanoma stem cells. a Heatmap of the differential mRNAs bound to the hnRNP A2B1 protein in melanoma stem cells and non-stem cells. b The expression profiles of mRNAs bound to the hnRNP A2B1 protein in melanoma cancer cells and non-stem cells. Quantitative real-time PCR was performed to detect the mRNA level (*p < 0.05, **p < 0.01). The experiments were biologically repeated three times. c KEGG classification of the differentially expressed genes. d Impact of hnRNP A2B1 silencing on cell viability and apoptosis of melanoma non-stem cells. Melanoma non-stem cells were transfected with hnRNP A2B1-siRNA or hnRNP A2B1-siRNA-scrambled. At 36 h after transfection, the cell viability and apoptosis of melanoma non-stem cells were examined. No statistical significance of difference between treatments was observed. e The sequence analysis of RIP in melanoma stem cells and non-stem cells using hnRNP A2B1 antibody. Black boxes and lines represented exons and introns, respectively. f The splicing mediated by hnRNP A2B1 in melanoma stem cells. Melanoma stem cells were transfected with hnRNP A2B1-siRNA or hnRNP A2B1-siRNA-scrambled. At 36 h after transfection, total RNAs were extracted and then subjected to semi-quantitative RT-PCR using intron-specific primers. β-tubulin was used as a control. g Influence of hnRNP A2B1 silencing on the expression levels of TPPP3, DOCK2, EIF3H, RNF128, DAPK1, and SYT7 in melanoma stem cells. At 36 h after transfection with hnRNP A2B1-siRNA or hnRNP A2B1-siRNA-scrambled, the expression levels of 6 proteins were examined using Western blot. β-tubulin was used as a control. The experiments were repeated three times. h Impact of hnRNP A2B1 overexpression on the expression levels of TPPP3, DOCK2, EIF3H, RNF128, DAPK1, and SYT7 in melanoma stem cells. At 48 h after the transfection of the recombinant plasmid expressing hnRNP A2B1, the expression levels of 6 proteins were detected using Western blot. The experiments were biologically repeated three times