Fig. 4

Universal verification of the single-vector dCasRx translation enhancement tool. A After constructing the single-vector CRISPR-dCasRx-eIF4GI tool targeting NIX (SV-NIX), the mRNA expression level of NIX was detected using qRT–PCR analysis. B The protein expression of NIX was verified using western blot and C the bar graph shows the relative expression level of NIX. D After constructing the single-vector CRISPR-dCasRx-eIF4GI tool targeting SIRT1 (SV-SIRT1), the mRNA expression level of SIRT1 was detected using qRT–PCR analysis. E The protein expression of SIRT1 was verified using western blot and F the bar graph shows the relative expression level of SIRT1. G After transfecting the plasmids containing the weak promoter with EGFP (pZDonor-PGK-EGFP) or negative control into HK2 cells for 8 h, respectively, the HK-2 cells were observed under an inverted fluorescence microscope and images were taken under dark field (magnification ×100). H After stably transfecting dCasRx-eIF4GI-EGFP into HK-2 cells, the plasmids containing the weak promoter with EGFP (pZDonor-PGK-EGFP) or negative control into HK2 cells for 8 h, respectively, the HK-2 cells were observed under an inverted fluorescence microscope and images were taken under dark field (magnification ×100). Data are presented as the means ± SEM from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and ns, no significant difference, represents P > 0.05