Fig 5. Targeting of the NEMO exon 1B core promoter with CRISPR/dCas9 reduced NEMO expression in 293T cells.

(A) 293T cells were infected with a LentiCRISPR2.0-dCas9 construct containing exon 1B gRNA, and transduced cells were selected with puromycin. Western blotting for NEMO was then performed on control cells (positive control 293T and 50% of the amount of 293T cell lysate, or negative control clone 1.1 cells) or dCas9-infected pools (dP) or single cell clones (d1, d2, d3). (B) The indicated cell extracts were analyzed by Western blotting for G6PD and Cas9/dCas9. In both cases, Ponceau staining of the filters was done to ensure approximately equal loading of total protein.