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Fig. 4 In vitro antitumor effects of AKPC-siYT on 2D cells. a, RT-PCR results after codelivery of siYAP and siTAZ to HCC38 cells. PBS (negative control); Lipo-siYAP, -siTAZ, -siYT (positive control): lipofectamine containing siYAP, siTAZ, siYAP, and a siTAZ mixture; MC3-siYAP, -siTAZ, -siYT: MC3-LNP containing YAP siRNA, TAZ siRNA, and a YAP and TAZ siRNA mixture; AKPC-siYAP, -siTAZ, -siYT: AKPC-LNPs containing YAP siRNA, TAZ siRNA, a YAP and TAZ siRNA mixture. The concentrations of siRNAs were constant for all conditions (siRNA, 2 μg/mL). b, RT-PCR results of the downstream gene after codelivery of siYAP and siTAZ to HCC38 cells. Two-way ANOVA was used to determine the significance of the comparisons of data indicated in a, b (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). In all panels, error bars represent mean ± s.d. (n = 3) c, Annexin V/PI staining of HCC38 cells after treatments with PBS and LNPs. d, Cell viability measurements by WST-1 in HCC38 cells after treatments with LNPs. Ordinary one-way ANOVA was used to determine the significance of the comparisons of data (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). In all panels, error bars represent mean ± s.d. (n = 3). e, 1000 cells were seeded as single cells in six-well plate, cultured continuously for 12 days, and treated with LNPs on days 1, 4, and 8 (siRNA, 2 μg/mL). Crystal Violet was used to stain cells and count the number of cell colonies on day 12, indicating the proliferation capacity of the cells.