Fig. 4: Expression of AAV-dSaCas9-KRAB-MeCP2(TRD) vectors in the reporter-HEK293T and HEPG2 cells.

a The samples were harvested 1-,2-and 3-weeks-post-transduction, and the luciferase assay was performed. Weeks 1–3; Lanes 1,4,7: Naïve cells. Lanes 2,5,8: AAV-dSaCas9-/no-gRNA. Lanes 3,6,9: AAV-dSaCas9-KRAB-MeCP2(TRD)/gRNA1. The experiment was done in quadruplicates. The statistical analysis, as above. b dGFP-expression. The samples were harvested 1-,2-,3- and 4 weeks-post-transduction, and the Western Blot was performed using anti-GFP-Ab. Weeks 1–4: Lanes 1,7,13,19 Untransduced cells; Lanes 2,8,14,20 AAV-dSaCas9-/no-gRNA; Lanes 3,9,15,21 AAV-no-Sp1/NF-kB; dSaCas9-KRAB-MeCP2/gRNA1; Lanes 4,10,16,22 AAV-no-Sp1/NF-kB-dSaCas9-KRAB-MeCP2/gRNA1; Lanes 5,11,17,23 AAV-dSaCas9- Sp1/NF-kB/no-gRNA. Lanes 6,12,18,24 AAV-dSaCas9- Sp1/NF-kB-KRAB-MeCP2/gRNA1. One-sample T-tests were carried out across normalized data, determining significant differences in means compared to the theoretical mean of 1. The values of Lanes 1,7,13,19 were normalized to equal the theoretical mean of 1. ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01. ***P ≤ 0.001, ****P ≤ 0.0001. The densitometry was measured using ImageJ software. c The samples were harvested 1-,2-,3- and 4 weeks-post-transduction, and the Western Blot was performed using anti-Pcsk9-Ab. Weeks 1–4; Lane 1,7,13,19. Untransduced HEPG2 cells. Lane 2,8,14,20. No-KRAB-MeCP2/no-gRNA. Lane 3,9,15,21. gRNA1 targeting Psck9 vector. Lane 4,10,16,22. gRNA2 targeting Pcsk9 vector. Lane 5,11,17,23. gRNA3 targeting Psck9 vector. The statistical analysis was done as above. The densitometry was measured using ImageJ.