Fig. 1.

CRISPR/Cas9-mediated knockout of p22phox in Nox4/tetNox4-HEK293 cells. A, Schematic representation of the designed guide RNAs (gRNA) targeting 5′UTR (untranslated region; 1&7, green), exon 1 (2&3, red) or intron 1 (4–6, blue) of the p22phox gene (CYBA; on chromosome 16). Image taken from UCSC genome browser (http://genome-euro.ucsc.edu/index.html) is modified. B, Used combinations (Combi.) of different gRNAs A-G for transfection of HEK293 cells. C, D, Representative Western blot for p22phox, Nox4 and β-Actin protein expression in empty vector control cells (Vec) and after CRISPR/Cas9-mediated p22phox knockout with different combinations or single gRNAs as indicated and defined in B. Every combination (combi.) was tested in Nox4-HEK293 (C) or tetNox4-HEK293 cells (D). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article).