Fig. 5

Role of hnRNP A2B1 in tumorigenesis of melanoma stem cells in vivo. a A flow diagram of the in vivo experiments. b Effects of hnRNP A2B1 knockdown on tumor growth in nude mice injected with melanoma stem cells (A375 or MDA-MB-435). The melanoma stem cells were transfected with hnRNP A2B1-shRNA or hnRNP A2B1-shRNA-scrambled. Six weeks later, the nude mice were sacrificed and the solid tumors were observed. c Influence of hnRNP A2B1 silencing on tumor volume. The tumor volume of mice injected with melanoma stem cells that were transfected with hnRNP A2B1-shRNA or hnRNP A2B1-shRNA-scrambled was examined every week. The mean of 5 mice was indicated (**p < 0.01). d Impact of hnRNP A2B1 knockdown on tumor weight in nude mice. The solid tumors from nude mice were taken out and weighed (**p < 0.01). e The expressions of stemness genes in solid tumors. Western blot was used to examine the expression levels of stemness genes in the solid tumors of mice injected with melanoma stem cells that were transfected with hnRNP A2B1-shRNA or hnRNP A2B1-shRNA-scrambled. β-tubulin was used as a control. f Expression level of hnRNP A2B1 in solid tumors of mice. Total RNAs extracted from the solid tumors of hnRNP A2B1-shRNA- or hnRNP A2B1-shRNA-scrambled-transfected mice were analyzed using quantitative real-time PCR (**p < 0.01). g Protein level of hnRNP A2B1 in the solid tumors of mice injected with melanoma stem cells transfected with hnRNP A2B1-shRNA or hnRNP A2B1-shRNA-scrambled. Western blot analysis was conducted to examine the hnRNP A2B1 protein level. β-tubulin was used as a control. h Effects of hnRNP A2B1 silencing on its targets’ expression levels in solid tumors. Western blot analysis was conducted to examine the protein levels of hnRNP A2B1’s targets in the solid tumors of mice treated with hnRNP A2B1-shRNA or hnRNP A2B1-shRNA-scrambled. β-tubulin was used as a control. i Immunohistochemical analysis of solid tumors. Solid tumors were analyzed by immunohistochemistry using the antibody against ki67 or Caspase 3 (brown). The nuclei were stained by DAPI (blue). Scale bar, 50 μm