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Fig. 5 Effects of SHK on the growth of leukemia cells in zebrafish xenografts. (A) Schematic illustration depicting the experimental protocol. Kasumi-1 cells labeled with CM-DiI fluorescent dye were injected into the yolk sac of zebrafish larvae at 48 hpf using microinjection techniques, as described in the Materials and Methods section. (B) The xenografts were treated with either vehicle (DMSO) or SHK at concentrations of 0.2 ?M for 24?72 h in a 34°C incubator. The fluorescence of Kasumi-1 cells within the larvae was observed under an inverted fluorescence microscope, and representative images were shown. (C) Human-specific GAPDH mRNA and (D) human-specific ?-actin mRNA were measured using RT?qPCR analysis. The levels of human-specific GAPDH and ?-actin mRNA were normalized to the zebrafish-specific ?-actin mRNA in the same sample. Data are presented as mean ± SD. **p < 0.01 indicates significant differences compared to the vehicle-treated groups.