Figure S2.

Silencing of the AAVS1^GFP/TetO7 Reporter Is Effective in K-562 Cells but Not in B-Lymphoblastoid Cells, Related to Figure 2

(A) Left: time-course flow cytometry analysis of AAVS1^GFP/TetO7 K-562 cell Clone #10 upon transfection with plasmids encoding for the indicated ETRs. Data show percentage of eGFP-negative cells (mean ± SEM; n = 3 independent transfections for each treatment condition). Right: representative dot plots of the indicated treatments at termination of the experiment.

(B) Fold change in the expression levels of the indicated genes of eGFP-negative cells sorted from the double ETR transfected conditions of (A). The expression level of each gene was normalized to B2M and represented as fold change over matched AAVS1^GFP/TetO7 untreated clone. Data are represented as mean ± SEM (n = 3 analyses on sorted cells from 3 independent transfections; statistical analysis by unpaired Student’s t test).

(C) Generation of the LV^TetO7/GFP cell lines. Representative dot plots showing transduction of K-562 cells (top), B-lymphoblastoid cells (middle) and NIH 3T3 cells (bottom) with LV^TetO7/GFP and relative sorting strategies to generate LV^TetO7/GFP reporter cell lines (dot plot on the right).

(D) Percentage of eGFP-positive cells at day 3 after the indicated treatments (mean ± SEM; n = 3 independent treatments; statistical analysis by unpaired Student’s t test).

(E) Time-course flow cytometry analysis of AAVS1^GFP/TetO7 B-lymphoblastoid cells upon transfection with in vitro transcribed mRNAs encoding for the indicated ETRs. Data show percentage of eGFP-negative cells (mean ± SEM; n = 3 independent transfections for each treatment condition).